Nucleic Acid Therapy Purity Methods by Capillary Gel ElectrophoresisCovance
The number of FDA-approved nucleic acid therapies including oligonucleotides and gene therapies has more than doubled since 2013. Safety, identity, purity and quality is of critical importance, so the characterization and routine testing of manufactured NAT drug substances and drug products is increasingly significant for public safety.
Recombinant Fab fragments specific for the pfHRPII and pfHSP72 : implications for malaria diagnosis - Parole de junior de la 7e édition du Cours international « Atelier Paludisme » - Ravaoarisoa Elisabeth - Madagascar - elisa@pasteur.mg
Nucleic Acid Therapy Purity Methods by Capillary Gel ElectrophoresisCovance
The number of FDA-approved nucleic acid therapies including oligonucleotides and gene therapies has more than doubled since 2013. Safety, identity, purity and quality is of critical importance, so the characterization and routine testing of manufactured NAT drug substances and drug products is increasingly significant for public safety.
Recombinant Fab fragments specific for the pfHRPII and pfHSP72 : implications for malaria diagnosis - Parole de junior de la 7e édition du Cours international « Atelier Paludisme » - Ravaoarisoa Elisabeth - Madagascar - elisa@pasteur.mg
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...AvactaLifeSciences
Avacta Life Sciences Exhibits Affimers at Global Protein Engineering Summit
Avacta Life Sciences exhibited recently at the Global Protein Engineering Summit ("PEGS") where it presented its Affimer technology.
You can read more about Affimer technology here http://www.avactalifesciences.com
PEGS is considered to be the essential protein engineering meeting where commercial and academic progress in protein engineering is showcased and this year it attracted over 1800 delegates from across the globe to Boston. Avacta Life Sciences presented its Affimer technology for the first time at a PEGS meeting with technical exhibits and a presentation by the CSO, Paul Ko Ferrigno, entitled "Biological Recognition: Beyond the Antibody."*
The exhibition booth was busy with over 80 delegates talking to the Avacta Life Science management team over the four days of the summit. The feedback on the Affimer technology was very positive, in particular, the short development times and excellent stability were highlighted by delegates as key advantages of Affimers over antibodies. There was also a strong interest in Affimers from the management of companies developing biological therapeutics who were keen to learn more about the potential of Affimers as novel therapeutics.
In addition, several companies were interested in the use of Affimers as an alternative to antibodies in diagnostic devices, mainly because they could generate binders against new biomarkers much more quickly and evaluate them in higher numbers.
The benefits of Affimer microarrays for biomarker discovery also resonated with diagnostic developers who appreciated the advantage of being able to evaluate significantly larger numbers of potential biomarkers more cost and time effectively than by mass spectrometry. The potential of the arrays for multiplexed solutions for clinical diagnosis and monitoring during drug trials was also something that generated interest amongst those delegates.
Matt Johnson, Chief Technical Officer of Avacta Life Sciences commented: "It was great to experience face to face the level of interest in Affimers. The majority of people I spoke to were either having problems raising antibodies to their target of interest or just couldn't use antibodies because of the type of assays they wanted to perform. Many of the presentations focused around the use of antibody fragments for intra-cellular studies which is a rapidly growing area that holds great interest for drug and diagnostics developers. It is an area where there are clear advantages for Affimers over antibody fragments which don't behave well in the cytoplasm.
"The general enthusiasm around Affimers was very encouraging and the amount of interest generated by the potential of Affimers as therapeutics and by the Affimer arrays for biomarker discovery only reinforces my excitement around this new technology."
Native peptides can be regarded as surrogate markers for protease activity in biological samples. Analysis of peptides by peptidomics allows to monitor protease activity in vivo and to describe the influence of protease inhibition. To elucidate the potential of peptides as markers for in vivo protease inhibition we analyzed plasma samples from animals treated with either the indirect FXa inhibitor FONDAPARINUX or the dipeptidylpeptidase IV inhibitor AB192. Signals correlating with the treatment were subsequently identified and assessed with respect to protease-dependent consensus cleavage motifs and occurrence of downstream targets. It could be shown that regulated peptides were either substrates, products or downstream targets of the inhibited protease. The results from the present study demonstrate that the in vivo analysis of peptides by peptidomics has the potential to broaden the knowledge of inhibitor related effects in vivo and that this method may pave the way to develop predictive biomarkers.
A Turn-Key Flow-Through-Mode Purification Process to improve Quality and Safe...Merck Life Sciences
In this webinar, you will learn:
Intensified plasma Immunoglobulin purifications
Scalable process development with latest technologies
Improved safety and quality of plasma IgG meeting required quality attributes
Detailed description:
Plasma-derived immunoglobulins (IgG) are essential medicines that are in worldwide shortage. How to develop optimized processing steps for robust and efficient manufacturing has been a constant goal, to make the most out of the precious plasma raw material.
In this study, we present a worse-case equivalent of plasma intermediate, explore various process steps along the fractionation flow, including flow-through-mode chromatography, affinity chromatography, virus inactivation steps and removal of solvent/detergent, single-pass TFF (SPTFF), clarification, and aseptic filtration, to establish a robust, easy-to-operate, readily scalable plasma IgG process with over 99% purity, depletion of IgA, isoagglutinin, and thrombogenic markers, meeting the commonly required 20% concentration for subcutaneous IgG infusion. Such solutions would be appropriate for various IgG intermediates which help to improve the global supply of immunoglobulins.
Development, safety and efficacy analysis of liquid state rabiesBalaganesh Kuruba
Rabies is a highly fatal epidemic disease in the world with high mortality rate in the infected individuals. According to the survey conducted by WHO across different parts of the globe, every year 50000 people die because of Rabies. And most of the vaccines are produced as solid-state vaccines.
Before formulation the purified PV 11 derived concentrated, infected and chromatographically purified rabies antigens are checked for their efficiency, potency by invitro methods.
Four different combinations of stabilizers, additives and adjuvants are blended with rabies antigen. Those are labelled as TCARLV-A, TCARLV-B, TCARLV-C, TCARLV-D. And find estimate the constituents in single Human dose.
The CiToxLAB Group offers a comprehensive range of preclinical services and specialty safety evaluation services to meet the needs of pharmaceutical, biotechnology and chemical companies worldwide.
Our four CiToxLAB international facilities in France, Canada, Denmark and Hungary carry out studies in general and reproductive toxicology, carcinogenicity, bioanalysis, immunology and safety pharmacology. The group has special expertise in areas such as inhalation or intra-nasal toxicology, radiation safety (ARS), NHPs and minipigs. Environmental studies are a further specialty including ecotoxicology and those related to REACH regulations.
Together with partners such as Atlanbio (St Nazaire, France) Stemina (Madison, USA) and Biomodels (Boston, USA), CiToxLAB also provides services such as clinical bioanalysis, embryonic stem cell biomarker discovery and customized preclinical efficacy models.
CiToxLAB now offers flexibility, direct contact to scientists, easy access to management and local, smart-sized facilities. Aggressive scheduling, increased size and capacity, turnkey solutions of global packages supported by project managers and broader geographic proximity are core values of CITOXLAB.
Contact our team of experts: www.citoxlab.com
Hot-start DNA polymerases are commonly used in PCR for genotyping, sequencing, molecular diagnostics, and high-throughput applications. In this presentation, PCR performance of Invitrogen™ Platinum II Taq Hot-Start DNA Polymerase and Invitrogen™ AccuPrime Taq DNA Polymerase is compared in the following areas:
• PCR run time for targets of different lengths
• Amplification of AT-rich and GC-rich sequences
• Tolerance to PCR inhibitors
• Sensitivity in target detection
• Universal protocol for PCR targets of different lengths
• Multiplex PCR of 15 targets
• Product format for direct gel loading
Request a sample of Platinum II Taq enzyme at http://bit.ly/2M4U9cw
Find other PCR enzymes at http://bit.ly/2JIPrzj
Learn more about PCR at http://bit.ly/2y2aSVo
#PCR #PCREducation #Invitrogen #InvitrogenSchoolofMolBio
Webinar: Evaluating Viral Clearance for Continuous ProcessesMilliporeSigma
Participate in the interactive webinar now: http://bit.ly/ViralClearanceWebinar
Is viral clearance a hurdle to implementing continuous processing? We’ll share virus spiking alternatives that may pave the way for effectively evaluating viral clearance by chromatography steps in a continuous process.
Explore our webinar library: www.emdmillipore.com/webinars
Beta-actin (human gene and protein symbol ACTB/ACTB) is one of six different actin isoforms which have been identified in humans. This is one of the two nonmuscle cytoskeletal actins. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/actin-b-antibody?filter_name=STJ91464
A polyhistidine-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, His6 tag and by the trademarked name His-tag. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/his-tag-antibody-p-95792?filter_name=STJ90106
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...AvactaLifeSciences
Avacta Life Sciences Exhibits Affimers at Global Protein Engineering Summit
Avacta Life Sciences exhibited recently at the Global Protein Engineering Summit ("PEGS") where it presented its Affimer technology.
You can read more about Affimer technology here http://www.avactalifesciences.com
PEGS is considered to be the essential protein engineering meeting where commercial and academic progress in protein engineering is showcased and this year it attracted over 1800 delegates from across the globe to Boston. Avacta Life Sciences presented its Affimer technology for the first time at a PEGS meeting with technical exhibits and a presentation by the CSO, Paul Ko Ferrigno, entitled "Biological Recognition: Beyond the Antibody."*
The exhibition booth was busy with over 80 delegates talking to the Avacta Life Science management team over the four days of the summit. The feedback on the Affimer technology was very positive, in particular, the short development times and excellent stability were highlighted by delegates as key advantages of Affimers over antibodies. There was also a strong interest in Affimers from the management of companies developing biological therapeutics who were keen to learn more about the potential of Affimers as novel therapeutics.
In addition, several companies were interested in the use of Affimers as an alternative to antibodies in diagnostic devices, mainly because they could generate binders against new biomarkers much more quickly and evaluate them in higher numbers.
The benefits of Affimer microarrays for biomarker discovery also resonated with diagnostic developers who appreciated the advantage of being able to evaluate significantly larger numbers of potential biomarkers more cost and time effectively than by mass spectrometry. The potential of the arrays for multiplexed solutions for clinical diagnosis and monitoring during drug trials was also something that generated interest amongst those delegates.
Matt Johnson, Chief Technical Officer of Avacta Life Sciences commented: "It was great to experience face to face the level of interest in Affimers. The majority of people I spoke to were either having problems raising antibodies to their target of interest or just couldn't use antibodies because of the type of assays they wanted to perform. Many of the presentations focused around the use of antibody fragments for intra-cellular studies which is a rapidly growing area that holds great interest for drug and diagnostics developers. It is an area where there are clear advantages for Affimers over antibody fragments which don't behave well in the cytoplasm.
"The general enthusiasm around Affimers was very encouraging and the amount of interest generated by the potential of Affimers as therapeutics and by the Affimer arrays for biomarker discovery only reinforces my excitement around this new technology."
Native peptides can be regarded as surrogate markers for protease activity in biological samples. Analysis of peptides by peptidomics allows to monitor protease activity in vivo and to describe the influence of protease inhibition. To elucidate the potential of peptides as markers for in vivo protease inhibition we analyzed plasma samples from animals treated with either the indirect FXa inhibitor FONDAPARINUX or the dipeptidylpeptidase IV inhibitor AB192. Signals correlating with the treatment were subsequently identified and assessed with respect to protease-dependent consensus cleavage motifs and occurrence of downstream targets. It could be shown that regulated peptides were either substrates, products or downstream targets of the inhibited protease. The results from the present study demonstrate that the in vivo analysis of peptides by peptidomics has the potential to broaden the knowledge of inhibitor related effects in vivo and that this method may pave the way to develop predictive biomarkers.
A Turn-Key Flow-Through-Mode Purification Process to improve Quality and Safe...Merck Life Sciences
In this webinar, you will learn:
Intensified plasma Immunoglobulin purifications
Scalable process development with latest technologies
Improved safety and quality of plasma IgG meeting required quality attributes
Detailed description:
Plasma-derived immunoglobulins (IgG) are essential medicines that are in worldwide shortage. How to develop optimized processing steps for robust and efficient manufacturing has been a constant goal, to make the most out of the precious plasma raw material.
In this study, we present a worse-case equivalent of plasma intermediate, explore various process steps along the fractionation flow, including flow-through-mode chromatography, affinity chromatography, virus inactivation steps and removal of solvent/detergent, single-pass TFF (SPTFF), clarification, and aseptic filtration, to establish a robust, easy-to-operate, readily scalable plasma IgG process with over 99% purity, depletion of IgA, isoagglutinin, and thrombogenic markers, meeting the commonly required 20% concentration for subcutaneous IgG infusion. Such solutions would be appropriate for various IgG intermediates which help to improve the global supply of immunoglobulins.
Development, safety and efficacy analysis of liquid state rabiesBalaganesh Kuruba
Rabies is a highly fatal epidemic disease in the world with high mortality rate in the infected individuals. According to the survey conducted by WHO across different parts of the globe, every year 50000 people die because of Rabies. And most of the vaccines are produced as solid-state vaccines.
Before formulation the purified PV 11 derived concentrated, infected and chromatographically purified rabies antigens are checked for their efficiency, potency by invitro methods.
Four different combinations of stabilizers, additives and adjuvants are blended with rabies antigen. Those are labelled as TCARLV-A, TCARLV-B, TCARLV-C, TCARLV-D. And find estimate the constituents in single Human dose.
The CiToxLAB Group offers a comprehensive range of preclinical services and specialty safety evaluation services to meet the needs of pharmaceutical, biotechnology and chemical companies worldwide.
Our four CiToxLAB international facilities in France, Canada, Denmark and Hungary carry out studies in general and reproductive toxicology, carcinogenicity, bioanalysis, immunology and safety pharmacology. The group has special expertise in areas such as inhalation or intra-nasal toxicology, radiation safety (ARS), NHPs and minipigs. Environmental studies are a further specialty including ecotoxicology and those related to REACH regulations.
Together with partners such as Atlanbio (St Nazaire, France) Stemina (Madison, USA) and Biomodels (Boston, USA), CiToxLAB also provides services such as clinical bioanalysis, embryonic stem cell biomarker discovery and customized preclinical efficacy models.
CiToxLAB now offers flexibility, direct contact to scientists, easy access to management and local, smart-sized facilities. Aggressive scheduling, increased size and capacity, turnkey solutions of global packages supported by project managers and broader geographic proximity are core values of CITOXLAB.
Contact our team of experts: www.citoxlab.com
Hot-start DNA polymerases are commonly used in PCR for genotyping, sequencing, molecular diagnostics, and high-throughput applications. In this presentation, PCR performance of Invitrogen™ Platinum II Taq Hot-Start DNA Polymerase and Invitrogen™ AccuPrime Taq DNA Polymerase is compared in the following areas:
• PCR run time for targets of different lengths
• Amplification of AT-rich and GC-rich sequences
• Tolerance to PCR inhibitors
• Sensitivity in target detection
• Universal protocol for PCR targets of different lengths
• Multiplex PCR of 15 targets
• Product format for direct gel loading
Request a sample of Platinum II Taq enzyme at http://bit.ly/2M4U9cw
Find other PCR enzymes at http://bit.ly/2JIPrzj
Learn more about PCR at http://bit.ly/2y2aSVo
#PCR #PCREducation #Invitrogen #InvitrogenSchoolofMolBio
Webinar: Evaluating Viral Clearance for Continuous ProcessesMilliporeSigma
Participate in the interactive webinar now: http://bit.ly/ViralClearanceWebinar
Is viral clearance a hurdle to implementing continuous processing? We’ll share virus spiking alternatives that may pave the way for effectively evaluating viral clearance by chromatography steps in a continuous process.
Explore our webinar library: www.emdmillipore.com/webinars
Beta-actin (human gene and protein symbol ACTB/ACTB) is one of six different actin isoforms which have been identified in humans. This is one of the two nonmuscle cytoskeletal actins. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/actin-b-antibody?filter_name=STJ91464
A polyhistidine-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, His6 tag and by the trademarked name His-tag. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/his-tag-antibody-p-95792?filter_name=STJ90106
Cdc2 Polyclonal Antibody plays a key role in the control of the eukaryotic cell cycle by modulating the centrosome cycle as well as mitotic onset; promotes G2-M transition, and regulates G1 progress and G1-S transition via association with multiple interphase cyclins.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/cdc2-antibody-p-91597?filter_name=STJ92154
NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/nfkb-p65-antibody-p-93367?filter_name=STJ94468
Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals. MTOR directly or indirectly regulates the phosphorylation of at least 800 proteins. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2). Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B, and the inhibitor of translation initiation PDCD4. Stimulates the pyrimidine biosynthesis pathway, both by acute regulation through RPS6KB1-mediated phosphorylation of the biosynthetic enzyme CAD, and delayed regulation, through transcriptional enhancement of the pentose phosphate pathway which produces 5-phosphoribosyl-1-pyrophosphate (PRPP), an allosteric activator of CAD at a later step in synthesis, this function is dependent on the mTORC1 complex. Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 an RNA polymerase III-repressor.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/mtor-antibody-p-93228?filter_name=STJ94280
Component of the epithelial apical junction complex that may function as an homophilic cell adhesion molecule and is essential for tight junction integrity. Also involved in transepithelial migration of leukocytes through adhesive interactions with AMICA1/JAML a transmembrane protein of the plasma membrane of leukocytes. The interaction between both receptors also mediates the activation of gamma-delta T-cells, a subpopulation of T-cells residing in epithelia and involved in tissue homeostasis and repair. Upon epithelial CXADR-binding, AMICA1 induces downstream cell signaling events in gamma-delta T-cells through PI3-kinase and MAP kinases. It results in proliferation and production of cytokines and growth factors by T-cells that in turn stimulate epithelial tissues repair.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/car-antibody?filter_name=STJ91996
Activity of CDK2 is maximal during S phase and G2; activated by interaction with cyclin E during the early stages of DNA synthesis to permit G1-S transition, and subsequently activated by cyclin A2 (cyclin A1 in germ cells) during the late stages of DNA replication to drive the transition from S phase to mitosis, the G2 phase. EZH2 phosphorylation promotes H3K27me3 maintenance and epigenetic gene silencing. Phosphorylates CABLES1 (By similarity). Cyclin E/CDK2 prevents oxidative stress-mediated Ras-induced senescence by phosphorylating MYC. Involved in G1-S phase DNA damage checkpoint that prevents cells with damaged DNA from initiating mitosis; regulates homologous recombination-dependent repair by phosphorylating BRCA2, this phosphorylation is low in S phase when recombination is active, but increases as cells progress towards mitosis. In response to DNA damage, double-strand break repair by homologous recombination a reduction of CDK2-mediated BRCA2 phosphorylation. Phosphorylation of RB1 disturbs its interaction with E2F1. NPM1 phosphorylation by cyclin E/CDK2 promotes its dissociates from unduplicated centrosomes, thus initiating centrosome duplication. Cyclin E/CDK2-mediated phosphorylation of NPAT at G1-S transition and until prophase stimulates the NPAT-mediated activation of histone gene transcription during S phase. Required for vitamin D-mediated growth inhibition by being itself inactivated. Involved in the nitric oxide- (NO) mediated signaling in a nitrosylation/activation-dependent manner. USP37 is activated by phosphorylation and thus triggers G1-S transition.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/cdk2-antibody-p-91630?filter_name=STJ92197
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/gapdh-antibody-p-94882?filter_name=STJ96417
GSK3β Polyclonal Antibody is an active protein kinase that acts as a negative regulator in the hormonal control of glucose homeostasis, Wnt signaling and regulation of transcription factors and microtubules, by phosphorylating and inactivating glycogen synthase (GYS1 or GYS2), EIF2B, CTNNB1/beta-catenin, APC, AXIN1, DPYSL2/CRMP2, JUN, NFATC1/NFATC, MAPT/TAU and MACF1. Requires primed phosphorylation of the majority of its substrates.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/gsk3b-antibody-p-92568?filter_name=STJ93447
Western Blot Antibody Customer Review for Anti-Phospho-ALK (Y1507) Polyclonal...St John's Laboratory Ltd
Anaplastic lymphoma kinase (ALK) also known as ALK tyrosine kinase receptor or CD246 (cluster of differentiation 246) is an enzyme that in humans is encoded by the ALK gene.
ALK plays an important role in the development of the brain and exerts its effects on specific neurons in the nervous system. Transduces signals from ligands at the cell surface, through specific activation of the mitogen-activated protein kinase (MAPK) pathway. Phospho-ALK (Y1507) Polyclonal Antibody detects endogenous levels of ALK protein only when phosphorylated at Y1507.
Anti-Phospho-ALK - http://www.stjohnslabs.com/phospho-alk-y1507-antibody?filter_name=STJ90845
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.
To purchase this antibody use the following link: http://www.stjohnslabs.com/trimethyl-histone-h3-k56?filter_name=STJ29403
Fukutin-related protein (FKRP) is targeted to the medial Golgi apparatus and is necessary for post-translational modification of dystroglycan. Mutations in the gene encoding this protein have been associated with congenital muscular dystrophy, mental retardation, and cerebellar cysts. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has not been determined.
To purchase this antibody use the following link: http://www.stjohnslabs.com/fkrp-antibody-p-95246?filter_name=FKRP
Histones are nuclear proteins that form octameric structures which bind DNA to form units of chromatin called nucleosomes. The family of histones—H2A, H2B, H3, and H4—are key players in gene regulation. They undergo a number of post-translational modifications (PTM) in response to various stimuli, including phosphorylation on serine and threonine residues and methylation on lysine residues. PTMs produce configural changes in histone proteins that may induce nucleosome remodeling and expose or hide DNA sequences from transcriptional complexes. Histone H4 lysine 20 (H4K20) may undergo mono-, di-, or trimethylation, which is catalyzed by the methyltransferase PR-Set7 (Set8 or KMT5a). Methylated H4K20 plays a role in regulating DNA damage responses, mitosis, DNA replication, and gene expression. Trimethylation of H4K20 contributes to gene silencing, and is a mark of the repressive heterochromatin state.
To purchase this antibody use the following link: http://www.stjohnslabs.com/histone-h4-tri-methyl-lys20-antibody?filter_name=STJ97154
Tyrosine-protein kinase that plays an essential role as cell surface receptor for neuregulins and EGF family members and regulates development of the heart, the central nervous system and the mammary gland, gene transcription, cell proliferation, differentiation, migration and apoptosis. Required for normal cardiac muscle differentiation during embryonic development, and for postnatal cardiomyocyte proliferation. Required for normal development of the embryonic central nervous system, especially for normal neural crest cell migration and normal axon guidance. Required for mammary gland differentiation, induction of milk proteins and lactation. Acts as cell-surface receptor for the neuregulins NRG1, NRG2, NRG3 and NRG4 and the EGF family members BTC, EREG and HBEGF. Ligand binding triggers receptor dimerization and autophosphorylation at specific tyrosine residues that then serve as binding sites for scaffold proteins and effectors. Ligand specificity and signaling is modulated by alternative splicing, proteolytic processing, and by the formation of heterodimers with other ERBB family members, thereby creating multiple combinations of intracellular phosphotyrosines that trigger ligand- and context-specific cellular responses. Mediates phosphorylation of SHC1 and activation of the MAP kinases MAPK1/ERK2 and MAPK3/ERK1. Isoform JM-A CYT-1 and isoform JM-B CYT-1 phosphorylate PIK3R1, leading to the activation of phosphatidylinositol 3-kinase and AKT1 and protect cells against apoptosis. Isoform JM-A CYT-1 and isoform JM-B CYT-1 mediate reorganization of the actin cytoskeleton and promote cell migration in response to NRG1. Isoform JM-A CYT-2 and isoform JM-B CYT-2 lack the phosphotyrosine that mediates interaction with PIK3R1, and hence do not phosphorylate PIK3R1, do not protect cells against apoptosis, and do not promote reorganization of the actin cytoskeleton and cell migration. Proteolytic processing of isoform JM-A CYT-1 and isoform JM-A CYT-2 gives rise to the corresponding soluble intracellular domains (4ICD) that translocate to the nucleus, promote nuclear import of STAT5A, activation of STAT5A, mammary epithelium differentiation, cell proliferation and activation of gene expression. The ERBB4 soluble intracellular domains (4ICD) colocalize with STAT5A at the CSN2 promoter to regulate transcription of milk proteins during lactation. The ERBB4 soluble intracellular domains can also translocate to mitochondria and promote apoptosis.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/erbb4?filter_name=STJ27886
Equilibrative nucleoside transporter 1 (ENT1) is a protein that in humans is encoded by the SLC29A1 gene. Multiple alternatively spliced variants, encoding the same protein, have been found for this gene.
This gene is a member of the equilibrative nucleoside transporter family. The gene encodes a transmembrane glycoprotein that localizes to the plasma and mitochondrial membranes and mediates the cellular uptake of nucleosides from the surrounding medium. The protein is categorized as an equilibrative (as opposed to concentrative) transporter that is sensitive to inhibition by nitrobenzylmercaptopurine ribonucleoside (NBMPR). Nucleoside transporters are required for nucleotide synthesis in cells that lack de novo nucleoside synthesis pathways, and are also necessary for the uptake of cytotoxic nucleosides used for cancer and viral chemotherapies.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/ent1-antibody?filter_name=STJ96396
Tau are stabilising proteins for axonal microtubules within the Central Nervous System, and as such are also recognised as Mictrotubule-associated Protein (MAP). Tau is predominantly active in the distal portions of axons to carry out its function.
Vesicle-associated membrane protein-associated protein B (VAPB) is a type IV membrane protein found in plasma and intracellular vesicle membranes. The protein is found as a homodimer and as a heterodimer with VAPA. It also can interact with VAMP1 and VAMP2 and may be involved in vesicle trafficking. Like VAPA, VAPB binds to proteins that contain a FFAT motif. Considerable interest in VAPB has arisen because mutations in this protein are associated with rare, familial forms of Motor Neurone Disease (also called Amyotrophic Lateral Sclerosis and Lou Gehrig's disease).
To purchase this antibody use the following link: http://www.stjohnslabs.com/vapb-antibody-c-term-p-79486?filter_name=STJ47066
Hypoxia-inducible factor 1-alpha, also known as HIF-1α, is a subunit of a heterodimeric transcription factor hypoxia-inducible factor 1 (HIF-1) that is encoded by the HIF1α gene. It is a basic helix-loop-helix PAS domain containing protein, and is considered as the master transcriptional regulator of cellular and developmental response to hypoxia. The dysregulation and overexpression of HIF1α by either hypoxia or genetic alternations have been heavily implicated in cancer biology, as well as a number of other pathophysiologies, specifically in areas of vascularization and angiogenesis, energy metabolism, cell survival and tumour invasion.
To purchase this antibody use the following link: http://www.stjohnslabs.com/hif-1a-antibody-p-92614?filter_name=STJ93498
Receptor tyrosine kinase which binds promiscuously membrane-bound ephrin-A family ligands residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. The signaling pathway downstream of the receptor is referred to as forward signaling while the signaling pathway downstream of the ephrin ligand is referred to as reverse signaling. Activated by the ligand ephrin-A1/EFNA1 regulates migration, integrin-mediated adhesion, proliferation and differentiation of cells. Regulates cell adhesion and differentiation through DSG1/desmoglein-1 and inhibition of the ERK1/ERK2 (MAPK3/MAPK1, respectively) signaling pathway. May also participate in UV radiation-induced apoptosis and have a ligand-independent stimulatory effect on chemotactic cell migration. During development, may function in distinctive aspects of pattern formation and subsequently in development of several fetal tissues. Involved for instance in angiogenesis, in early hindbrain development and epithelial proliferation and branching morphogenesis during mammary gland development. Engaged by the ligand ephrin-A5/EFNA5 may regulate lens fiber cells shape and interactions and be important for lens transparency development and maintenance. With ephrin-A2/EFNA2 may play a role in bone remodeling through regulation of osteoclastogenesis and osteoblastogenesis.
To purchase this antibody, use the following link: http://www.stjohnslabs.com/epha2-antibody-p-96094?filter_name=STJ92940
Antibody Customer Review For Phospho-eIF2α (S51) Polyclonal Antibody (STJ90253) St John's Laboratory Ltd
eIF2 is an initiation factor found within eukaryotes. It mediates binding of tRNA to the ribosome in a GTP dependent manner. The alpha subunit is the main target for phosphorylation at Serine-51, and is considered the regulatory subunit of this trimeric (formed of eIF2a, eIF2b and eIF2y).
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Similar to St John's Laboratory Ltd Booklet 2015 (20)
G protein-coupled receptor that probably associates with the patched protein (PTCH) to transduce the hedgehog's proteins signal. Binding of sonic hedgehog (SHH) to its receptor patched is thought to prevent normal inhibition by patched of smoothened (SMO). Required for the accumulation of KIF7 and GLI3 in the cilia.
Anti-Smo antibody (STJ95710): http://www.stjohnslabs.com/smo-antibody-p-94371?filter_name=STJ95710
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Western Blot Customer Review Anti Glucocorticoid Receptor antibody (STJ97101)St John's Laboratory Ltd
Receptor for glucocorticoids (GC). Has a dual mode of action: as a transcription factor that binds to glucocorticoid response elements (GRE), both for nuclear and mitochondrial DNA, and as a modulator of other transcription factors. Affects inflammatory responses, cellular proliferation and differentiation in target tissues. Involved in chromatin remodeling . Plays a role in rapid mRNA degradation by binding to the 5' UTR of target mRNAs and interacting with PNRC2 in a ligand-dependent manner. Could act as a coactivator for STAT5-dependent transcription upon growth hormone (GH) stimulation and could reveal an essential role of hepatic GR in the control of body growth (By similarity). Has transcriptional activation and repression activity . Mediates glucocorticoid-induced apoptosis . Promotes accurate chromosome segregation during mitosis . May act as a tumor suppressor . May play a negative role in adipogenesis through the regulation of lipolytic and antilipogenic gene expression (By similarity). / Isoform Beta: Acts as a dominant negative inhibitor of isoform Alpha . Has intrinsic transcriptional activity independent of isoform Alpha when both isoforms are coexpressed.
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Anti glucocorticoid receptor antibody (STJ97101):
http://www.stjohnslabs.com/glucocorticoid-receptor-antibody-p-98736?filter_name=STJ97101
Western Blot Customer Review Anti-Phospho-Cofilin (S3) Antibody (STJ90230)St John's Laboratory Ltd
Binds to F-actin and exhibits pH-sensitive F-actin depolymerizing activity. Regulates actin cytoskeleton dynamics. Important for normal progress through mitosis and normal cytokinesis. Plays a role in the regulation of cell morphology and cytoskeletal organization. Required for the up-regulation of atypical chemokine receptor ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation.
Anti-Phospho-Cofilin (S3) -http://www.stjohnslabs.com/phospho-cofilin-s3-antibody
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
This June, Dr. Byron Baron from the University of Malta, Malta, is our Scientist of the Month! He's shared with us his research highlights, his current projects and some comments on the biotechnology industry.
Want to be our Scientist of the Month? Contact info@stjohnslabs.com
Downstream effector molecule involved in the transmission of signals from tyrosine kinase receptors and small GTPases to the actin cytoskeleton. Promotes formation of actin filaments. Part of the WAVE complex that regulates lamellipodia formation. The WAVE complex regulates actin filament reorganization via its interaction with the Arp2/3 complex.
Anti-WAVE2 -http://www.stjohnslabs.com/wave2-antibody
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Implicated in synaptic vesicle endocytosis. May recruit other proteins to membranes with high curvature.
Brain, mostly in frontal cortex. Expressed at high level in fetal cerebellum.
Anti-Endophilin I -http://www.stjohnslabs.com/endophilin-i-antibody
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Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain. TUBB3 plays a critical role in proper axon guidance and mantainance.
Anti-β-tubulin -http://www.stjohnslabs.com/b-tubulin-antibody-p-98672
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Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-|-AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex. / Strict requirement for Asp at position P1 and has a preferred cleavage sequence of (Leu/Asp/Val)-Glu-Thr-Asp-|-(Gly/Ser/Ala). / Inhibited by the effector protein NleF that is produced by pathogenic E.coli; this inhibits apoptosis.
Anti-Caspase-8-http://www.stjohnslabs.com/caspase-8-antibody-p-99045
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Tubulin is the major constituent of microtubules. The gamma chain is found at microtubule organizing centers (MTOC) such as the spindle poles or the centrosome. Pericentriolar matrix component that regulates alpha/beta chain minus-end nucleation, centrosome duplication and spindle formation.
Anti-Gamma Tubulin-http://www.stjohnslabs.com/gamma-tubulin-antibody
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Tubulin in molecular biology can refer either to the tubulin protein superfamily of globular proteins, or one of the member proteins of that superfamily. α- and β-tubulins polymerize into microtubules, a major component of the eukaryotic cytoskeleton. Microtubules function in many essential cellular processes, including mitosis. Tubulin-binding drugs kill cancerous cells by inhibiting microtubule dynamics, which are required for DNA segregation and therefore cell division. In eukaryotes there are six members of the tubulin superfamily, although not all are present in all species (see below). Both α and β tubulins have a mass of around 50 kDa and are thus in a similar range compared to actin with ~42 kDa. In contrast, tubulin polymers (microtubules) tend to be much bigger than actin filaments due to their cylindrical nature. Tubulin was long thought to be specific to eukaryotes. More recently, however, several prokaryotic proteins have been shown to be related to tubulin.
Anti-Epsilon Tubulin -http://www.stjohnslabs.com/epsilon-tubulin-antibody-2
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Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes) . Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation .
Anti-LC3A-http://www.stjohnslabs.com/lc3a-antibody
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Multifunctional transcription factor in ER stress response. Plays an essential role in the response to a wide variety of cell stresses and induces cell cycle arrest and apoptosis in response to ER stress. Plays a dual role both as an inhibitor of CCAAT/enhancer-binding protein (C/EBP) function and as an activator of other genes. Acts as a dominant-negative regulator of C/EBP-induced transcription: dimerizes with members of the C/EBP family, impairs their association with C/EBP binding sites in the promoter regions, and inhibits the expression of C/EBP regulated genes. Positively regulates the transcription of TRIB3, IL6, IL8, IL23, TNFRSF10B/DR5, PPP1R15A/GADD34, BBC3/PUMA, BCL2L11/BIM and ERO1L. Negatively regulates; expression of BCL2 and MYOD1, ATF4-dependent transcriptional activation of asparagine synthetase (ASNS), CEBPA-dependent transcriptional activation of hepcidin (HAMP) and CEBPB-mediated expression of peroxisome proliferator-activated receptor gamma (PPARG).
Anti-CHOP-http://www.stjohnslabs.com/chop-antibody-2
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Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process. Activated following phosphorylation by RIPK3, leading to homotrimerization, localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage. Does not have protein kinase activity.
Anti-phospho-MLKL (S358)-http://www.stjohnslabs.com/phospho-mlkl-s358-antibody-1
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Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. The MAPK/ERK cascade plays also a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. About 160 substrates have already been discovered for ERKs. Many of these substrates are localized in the nucleus, and seem to participate in the regulation of transcription upon stimulation.
Anti-ERK1-http://www.stjohnslabs.com/erk1-antibody-3
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Tyrosine-protein kinase that acts as a cell-surface receptor for PDGFA, PDGFB and PDGFC and plays an essential role in the regulation of embryonic development, cell proliferation, survival and chemotaxis. Depending on the context, promotes or inhibits cell proliferation and cell migration. Plays an important role in the differentiation of bone marrow-derived mesenchymal stem cells. Required for normal skeleton development and cephalic closure during embryonic development. Required for normal development of the mucosa lining the gastrointestinal tract, and for recruitment of mesenchymal cells and normal development of intestinal villi. Plays a role in cell migration and chemotaxis in wound healing. Plays a role in platelet activation, secretion of agonists from platelet granules, and in thrombin-induced platelet aggregation.
Anti-PDGFRα-http://www.stjohnslabs.com/pdgfra-antibody-2
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Thiol protease that cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes. Important for defense against pathogens. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Can also promote apoptosis. / Strict requirement for an Asp residue at position P1 and has a preferred cleavage sequence of Tyr-Val-Ala-Asp-|-. / Specifically inhibited by the cowpox virus Crma protein.
Anti-Caspase-1-http://www.stjohnslabs.com/caspase-1-antibody-1
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Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Involved in cell mobility and transcription regulation through protein-protein interactions. Can promote transcription activation through binding to APBB1-KAT5 and inhibits Notch signaling through interaction with Numb. Couples to apoptosis-inducing pathways such as those mediated by G(O) and JIP. Inhibits G(o) alpha ATPase activity (By similarity). Acts as a kinesin I membrane receptor, mediating the axonal transport of beta-secretase and presenilin 1. Involved in copper homeostasis/oxidative stress through copper ion reduction. In vitro, copper-metallated APP induces neuronal death directly or is potentiated through Cu2+-mediated low-density lipoprotein oxidation.
Anti-Amyloid-β-http://www.stjohnslabs.com/amyloid-v-antibody
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Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release .
Anti-Bcl-2-http://www.stjohnslabs.com/bcl-2-antibody-1
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Alpha-actin-2 also known as actin, aortic smooth muscle or alpha smooth muscle actin (α-SMA, SMactin, alpha-SM-actin, ASMA) is a protein that in humans is encoded by the ACTA2 gene located on 10q22-q24. Actin alpha 2, the human aortic smooth muscle actin gene, is one of six different actin isoforms which have been identified. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus. Alpha-smooth muscle actin (α-SMA) is commonly used as a marker of myofibroblast formation.
Anti-α-SMA -http://www.stjohnslabs.com/a-sma-antibody-1
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Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
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Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
3. Improving in quality and reproducibility
through antibody validation.
- St John’s Laboratory Team
4.
5. St John's Laboratory was awarded "Most
Impressive Startup" by a panel of judges from
F1000 Research. St John's Laboratory was
recognized for their antibody validation project.
6. At St John’s Laboratory we would like to increase the
transparency of our antibodies' quality, while also providing
sufficient background data on the product’s performance in
real-experimental conditions.
Through our Premier Partner Scheme (PPS) we have been
able to connect our customers with not only a product, but
also the opportunity to evaluate the product’s performance
before making a purchase.
Our customer’s validation review provide other end-users
valuable information on:
- New applications
- Optimal dilution conditions
- Protocols
- Images of our products at work
7. VALIDATION REPORT: Cdk2 Polyclonal Antibody (STJ92197)
BACKGROUND
Antibody: Cdk2 Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ92197
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were
target bands detected by STJ92197. Cdk2 Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: Actin-Beta Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
PROTOCOL
Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
Blocking: 1X TBST with 5% skimmed milk powder for 2 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:
10000 dilution ratio, for 1.5 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualiza-
tion time was identical for the same antigen).
Cdk2 Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug St J92197. Cdk2
Polyclonal
Antibody
1:1000
1:10000 32KD 60S
2 Jurkat lysate 10 ug 1:10000 32KD 60S
Figure 1: Lane1: Hela; Lane 2: Jurkat
8. VALIDATION REPORT: GSK3β Polyclonal Antibody (STJ93447)
BACKGROUND
Antibody: GSK3β Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ93447
Supplier: St John’s Laboratory
Experimental Notes: In Hela cell line, there was target band detected by STJ93447. GSK3β
Polyclonal Antibody. In A549 cell line, there was no target band detected by STJ93447. GSK3
β Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: Actin-Beta Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for GSK3β Polyclonal Antibody
GSK3β Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug St J93447. GSK3β
Polyclonal
Antibody
1:1000
1:10000 47KD 30S
2 A549 lysate 10 ug 1:10000 47KD 30S
Figure 1: Lane1: Hela; Lane 2: A549
9. VALIDATION REPORT: mTOR Polyclonal Antibody (STJ94280)
BACKGROUND
Antibody: mTOR Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ94280
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were target bands
detected by STJ94280. mTOR Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: mTOR Polyclonal Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for mTOR Polyclonal Antibody
mTOR Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug STJ94280. mTOR
Polyclonal
Antibody
1:1000
1:10000 288KD 35S
2 Jurkat lysate 10 ug 1:10000 288KD 35S
Figure 1: Lane1: Hela; Lane 2: Jurkat
10. VALIDATION REPORT: NFκB-p65 Polyclonal Antibody (STJ94468)
BACKGROUND
Antibody: NFκB-p65 Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ94468
Supplier: St John’s Laboratory
Experimental Notes: In Hela cell line, there was a target band detected by
STJ94468. NFκB-p65 Polyclonal Antibody. In A549 cell line, there were no
target bands detected by STJ94468. NFκB-p65 Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: NFκB-p65 Polyclonal Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for NFκB-p65 Polyclonal Antibody
NFκB-p65 Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug STJ94468.
NFκB-p65
Polyclonal
Antibody
1:1000
1:10000 65KD 30S
2 A549 lysate 10 ug 1:10000 65KD 30S
Figure 1: Lane1: Hela; Lane 2: Jurkat
11. VALIDATION REPORT: p53 Polyclonal Antibody (STJ94890)
BACKGROUND
Antibody: p53 Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ94890
Supplier: St John’s Laboratory
Experimental Notes: In Jurkat cell line, there was a target
band detected by STJ94890. p53 Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: p53 Polyclonal Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for p53 Polyclonal Antibody
p53 Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug STJ94890. p53
Polyclonal
Antibody
1:1000
1:10000 53KD 30S
2 Jurkat lysate 10 ug 1:10000 53KD 30S
Figure 1: Lane1: Hela; Lane 2: Jurkat
12. VALIDATION REPORT: PCNA Polyclonal Antibody (STJ94982)
BACKGROUND
Antibody: PCNA Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ94982
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were target
bands detected by STJ94982. PCNA Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: PCNA Polyclonal Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for p53 Polyclonal Antibody
PCNA Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug St J94982. PCNA
Polyclonal
Antibody
1:1000
1:10000 36KD 30S
2 Jurkat lysate 10 ug 1:10000 36KD 30S
Figure 1: Lane1: Hela; Lane 2: Jurkat
13. VALIDATION REPORT: GAPDH Polyclonal Antibody (STJ96417)
BACKGROUND
Antibody: GAPDH Polyclonal Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ96417
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were target
bands detected by STJ96417. GAPDH Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: GAPDH Polyclonal Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
Table: Validation Results for GAPDH Polyclonal Antibody
GAPDH Polyclonal Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug STJ96417. GAPDH
Polyclonal
Antibody
1:1000
1:10000 36KD 130S
2 Jurkat lysate 10 ug 1:10000 36KD 130S
Figure 1: Lane1: Hela; Lane 2: Jurkat
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15. VALIDATION REPORT: Actin-Beta Antibody (STJ91464)
BACKGROUND
Antibody: Actin-Beta Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ91464
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were
target bands detected by STJ91464.Actin-beta antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: Actin-Beta Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
PROTOCOL
Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
Blocking: 1X TBST with 5% skimmed milk powder for 2 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:
10000 dilution ratio, for 1.5 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualiza-
tion time was identical for the same antigen).
Actin-Beta Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug STJ91464.
Actin-beta
antibody
1:1000
1:10000 ~50KD 20S
2 Jurkat lysate 10 ug 1:10000 ~50KD 20S
Figure 1: Lane1: Hela; Lane 2: Jurkat
16. VALIDATION REPORT: Cdc2 Polyclonal Antibody (STJ92154)
BACKGROUND
Antibody: Cdc2 Antibody
Method of Validation: Western Blot
Validator: BPI
Supplier Catalog Number: STJ92154
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were
target bands detected by STJ92154. Cdc2 Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: Cdc2 Antibody (Dilution 1:1000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
PROTOCOL
Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
Blocking: 1X TBST with 5% skimmed milk powder for 2 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:
10000 dilution ratio, for 1.5 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualiza-
tion time was identical for the same antigen).
Cdc2 Antibody has been independently validated for western blot by BPI.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug St J92154. Cdc2
Polyclonal
Antibody
1:1000
1:10000 34KD 13S
2 Jurkat lysate 10 ug 1:10000 34KD 13S
Figure 1: Lane1: Hela; Lane 2: Jurkat
17. Figure 1: Western Blot for EGFR. Grey
arrowhead indicates the expected
molecular weight of ~170 kDa.PROTOCOL
Cell/tissue total protein lysates were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5
minutes prior to loading.
15 μg of boiled lysate were loaded and resolved on a 12% SDS-polyacrylamide gel.
The Precision Plus Protein™ All Blue Prestained Standards from BioRad (161-0373) were used as molecular mass markers.
Proteins were transferred onto nitrocellulose membrane by tank transfer and protein transfer was confirmed with Ponceau S
staining.
The immunoblot membrane was blocked in 2.5% skim milk and 1.5% BSA solution in TTBS at room temperature for 60 minutes.
The membrane was washed in TTBS twice for 5 minutes each.
The membrane was immersed with the protein side up in the antibody solution in TBS and incubated overnight at 4°C with gentle
agitation.
The membrane was rinsed twice with TTBS.
The membrane was washed in TTBS twice for 5 minutes each.
The membrane was washed in TTBS once for 15 minutes.
The membrane was incubated in the HRP-conjugated secondary antibody solution in TBS for 60 minutes at room temperature
with gentle agitation.
The membrane was rinsed twice with TTBS.
The membrane was washed in TTBS twice for 5 minutes each.
The membrane was washed in TTBS once for 15 minutes.
Signals were detected by chemiluminescence (ECL). The blot was scanned for 320 seconds.
The membrane was rinsed three times with TTBS.
Repeated Steps 4-15 with the loading control antibody and its matching secondary antibody.
VALIDATION REPORT: Epidermal Growth Factor Receptor (EGFR) antibody (STJ40568)
BACKGROUND
Antibody: Epidermal Growth Factor Receptor (EGFR) antibody
Method of Validation: Western Blot
Validator: Kinexus Bioinformatics Corporation
Supplier Catalog Number: STJ40568
Supplier: St John’s Laboratory
Experimental Notes: A strong band was observed in the positive control at the
expected size (~170 kDa) that is not observed in the negative control.
INDEPENDENT VALIDATION METHODS
Primary Antibody: Epidermal Growth Factor Receptor (EGFR) antibody (Dilution 1:1,000)
Loading Control Antibody: Beta-actin; supplier: Santa Cruz (Dilution 1:200)
Secondary Antibody: Sheep anti-mouse IgG-HRP; supplier: GE Healthcare (Dilution 1:10,000)
Epidermal Growth Factor Receptor (EGFR) antibody has been independently validated for
western blot by Kinexus Bioinformatics Corporation under the Science Exchange network.
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18. VALIDATION REPORT: FAAH Antibody (STJ23602)
BACKGROUND
Antibody: FAAH Antibody
Method of Validation: Immunofluorescence
Validator: Natalia Malek (PhD Student)
Supplier Catalog Number: STJ23602
Supplier: St John’s Laboratory
Experimental Notes: We completed evaluation of FAAH antibody. It worked really well,
so we decided to purchase a vial from St John’s Laboratory.
INDEPENDENT VALIDATION METHODS
Primary Antibody: FAAH antibody (Dilution 1:50)
PROTOCOL
Samples were frozen and stored at -80°C until sections were cut at 12 μm thick and collected onto gelatin-coated slides (Menzel
GmbH, Braunschweig, Germany).
For immunofluorescence labeling, serial sections of DRG were hydrated in PB buffer (pH=7.4) for 15 mins.
Cell membranes were permeabilised in PB containing 0.1% Triton X-100 and afterwards incubated for 1 h in 10% normal goat serum
(NGS, Jackson ImmunoResearch Laboratories, West Grove, USA) in PB containing 0.4% Triton X-100 (blocking solution).
Sections were incubated for 48 hrs at 4°C in a humid chamber with the respective primary antibodies (FAAH, St. John’s
laboratory; 1:50).
After three washes in PB, triple immunofluorescence was revealed by incubation at 4°C in for 24 hrs in the dark with the
appropriate fluorochrome-conjugated secondary antibody.
Sections were washed 3 times for 15 mins with PB and coverslipped with Aquatex mounting medium (Merck, Darmstadt,
Germany).
The sections processed for immunofluorescence were studied with a fluorescence microscope (DM RXA2) with confocal scanner
(TCS SL) (Leica Microsystems GmbH, Manheim, Germany) equipped with the following lasers: Ar 488, He-Ne 543, He-Ne 633.
FAAH antibody has been independently validated for immunofluorescence by PhD
student from Institute of Pharmacology Polish Academy of Sciences.
Figure 1: Immunofluorescence. Dorsal
root ganglia stained with FAAH antibody.
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19. PROTOCOL
Plating the cells:
200k of HEK293 cells were plated in each wells in a 24 well plate and cultured overnight in serum free media in an incubator at 37C
with 5% CO2.
Transfection
Cells were transfected with pLV-mCherry using Lipofectamine 2000 according to manufacturer's protocol and grown overnight
before fixing.
Fixing/permeablization/staining
Cells were fixed with 4% PFA for 15 min at RT.
Cells were permeablized with 0.1% Triton-X100 for 30 min.
Afterwards, cells were blocked using 1% BSA for 30 min.
Cells were incubated with mCherry antibody (1:200, 1 mg/mL) or isotype control (1:400, 2 mg/mL) at RT for 1 hr.
Cells were washed with PBS 3X at 5 min each
Cells were incubated with Goat anti-Mouse IgG(H+L) conjugated with HRP (1:200) for 30 min at RT.
Colormetric signal was developed using DAB solutions.
VALIDATION REPORT: mCherry Antibody (STJ34373)
BACKGROUND
Antibody: mCherry Antibody
Method of Validation: Immunohistochemistry
Validator: Microstem Inc
Supplier Catalog Number: STJ34373
Supplier: St John’s Laboratory
Experimental Notes: Strong signal was detected in the positive
control, but not in the negative control.
INDEPENDENT VALIDATION METHODS
Primary Antibody: mCherry Antibody
Isotype Control Antibody: mouse IgG1; supplier: CusaBio
Secondary Antibody: Goat anti-Mouse IgG(H+L) conjugated with HRP; supplier: KPL
mCherrry antibody has been independently validated for Immunohistochemistry by Microstem Inc.
Figure 1: HEK293 cell transfected with mCherry
vector stained with mCherry antibody developed
using IHC method.
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20. VALIDATION REPORT: CD133 Antibody (STJ20168)
BACKGROUND
Antibody: CD133 Antibody
Method of Validation: Immunohistochemistry
Supplier Catalog Number: STJ20168
Supplier: St John’s Laboratory
Experimental Notes: Very satisfied! Great communication with
seller. The antibody tested stained intensely at the membrane
as predicted. Compared to antibodies from other companies,
this is the best CD133 staining I have obtained so far.
INDEPENDENT VALIDATION METHODS
Primary Antibody: CD133 Antibody (Dilution: 1:200 in PBS
containing 0.1% porcine gelatin)
CD133 antibody has been independently validated for Immunohistochemistry
by Tago Santos from Health Sciences Research Center.
Figure 1: Immunocytochemistry on
human endothelial progenitor cells
isolated from stroke patients. Images
acquired by confocal microscopy.
VALIDATION REPORT: RARA Antibody (STJ26160)
BACKGROUND
Antibody: RARA Antibody
Method of Validation: Western Blot
Supplier Catalog Number: STJ26160
Supplier: St John’s Laboratory
Experimental Notes: The antibody tested gave a clear band
at the predicted molecular weight.
INDEPENDENT VALIDATION METHODS
Primary Antibody: RARA Antibody (Dilution: 1:1000 in TBS
containing 0.1% porcine gelatin)
RARA antibody has been independently validated for Western
Blot by Tago Santos from Health Sciences Research Center.
Figure 1: Western blotting on
subventricular zone neural stem
cells isolated from mice (primary
culture) using ECL substrate.
21. PROTOCOL
Immunoblotting
After running SDS-gel and transferring proteins to nitrocellulose membrane, such steps were performed:
Blocking with 1X PBS-T containing 5% nonfat dry milk with constant shacking for 1 hour.
Incubation with primary antibody (dilution 1:1000 in blocking buffer) overnight at 4°C.
Washing the membrane with PBS-T 3x10 min.
Incubation with HRP-conjugated secondary antibody for 1h in room temperature.
Washing the membrane with PBS-T 3x10 min.
Imaging of the membrane.
VALIDATION REPORT: LPCAT1 Antibody (STJ27005)
BACKGROUND
Antibody: LPCAT1 Antibody
Method of Validation: Western Blot
Supplier Catalog Number: STJ27005
Supplier: St John’s Laboratory
Experimental Notes: Product worked as expected.
INDEPENDENT VALIDATION METHODS
Primary Antibody: LPCAT1 Antibody (Dilution: 1:1000)
LPCAT1 antibody has been independently validated for Western Blot by Kristina
Klizaite from Life & Medical Sciences University of Bonn.
Figure 1: Western blot image of
mouse lung tissue.
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22. PROTOCOL
Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
Blocking: 1X TBST with 5% skimmed milk powder for 2 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:10000
dilution ratio, for 1.5 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualization
time was identical for the same antigen).
VALIDATION REPORT: HA-Tag Polyclonal Antibody (STJ90107)
BACKGROUND
Antibody: HA-Tag Antibody
Method of Validation: Western Blot
Supplier Catalog Number: STJ90107
Supplier: St John’s Laboratory
Experimental Notes: In Hela and Jurkat cell line, there were target bands
detected by STJ90107. HA-tag Polyclonal Antibody.
INDEPENDENT VALIDATION METHODS
Primary Antibody: HA-Tag Antibody (Dilution 1:10000)
Secondary Antibody: HRP goat anti-rabbit (provided by BPI)
Reference Antibody: Actin (provided by BPI)
HA-Tag antibody has been independently validated for Western Blot by BPI
Figure 1: Western blot;Hela and Jurkat
cell line target bands detected by HA
Tag antibody.
Gel electrophoresis infromation:10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information:0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen Loading amount Primary antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD Visualization time
1 Hela lysate 10 ug St J90107. HA-tag
Polyclonal
Antibody
1:1000
1:10000 68KD 34S
2 Jurkat lysate 10 ug 1:10000 68KD 34S
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