Customer Review For GAPDH Polyclonal Antibody (STJ96417)

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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

HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: GAPDH Polyclonal Antibody (STJ96417)

WHAT WAS USED?
Primary antibody: STJ96417. GAPDH Polyclonal
Antibody. 100µl
Provider: St John's Laboratory
Dilution ratio: 1:1000
Application: Western Blot.
Image: In HeLa and Jurkat cell line, there were target
bands detected by STJ96417 GAPDH Polyclonal
Antibody.

WHAT WAS THE PROTOCOL?
Treatment of materials: When cells were cultured to 70%-80% plating density,
lysis buffer with protease inhibitors (Sigma) and equal volume of 2X loading buffer
were added for 10 min water bath at 100˚C.
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).

WHAT WAS THE PROTOCOL?
Gel electrophoresis information：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 µg STJ96417.
GAPDH
Polyclonal
Antibody
1:1000
1:10000 36KD 130S
2 Jurkat lysate 10 µg 1:10000 36KD 130S

WHAT DID THE CUSTOMER THINK?
Testing results were provided by BPI laboratory. They
stated that “In HeLa and Jurkat cell line, there were target
bands detected by STJ96417 GAPDH Polyclonal
Antibody”.

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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

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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

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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

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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

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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

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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

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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

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Heat shock proteins, as a class, are among the most highly expressed cellular proteins across all species. As their name implies, heat shock proteins protect cells when stressed by elevated temperatures. They account for 1–2% of total protein in unstressed cells. However, when cells are heated, the fraction of heat shock proteins increases to 4–6% of cellular proteins. Hsp90 (heat shock protein 90) is a chaperone protein that assists other proteins to fold properly, stabilizes proteins against heat stress, and aids in protein degradation. It is one of the most common of the heat-related proteins. The "90" comes from the fact that it weighs roughly 90 kiloDaltons. A 90 kDa protein is considered fairly large for a non-fibrous protein. To purchase this antibody use the following link: http://www.stjohnslabs.com/hsp90a-antibody-p-92714?filter_name=HSP90A

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Constitutively 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. In skeletal muscle, contributes to insulin regulation of glycogen synthesis by phosphorylating and inhibiting GYS1 activity and hence glycogen synthesis. May also mediate the development of insulin resistance by regulating activation of transcription factors. Regulates protein synthesis by controlling the activity of initiation factor 2B (EIF2BE/EIF2B5) in the same manner as glycogen synthase. In Wnt signaling, GSK3B forms a multimeric complex with APC, AXIN1 and CTNNB1/beta-catenin and phosphorylates the N-terminus of CTNNB1 leading to its degradation mediated by ubiquitin/proteasomes. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Anti-GSK3β-http://www.stjohnslabs.com/gsk3b-antibody-p-92568 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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PIWIL4 belongs to the Argonaute family of proteins, which function in development and maintenance of germline stem cells. Plays a central role during spermatogenesis by repressing transposable elements and prevent their mobilization, which is essential for the germline integrity. Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and govern the methylation and subsequent repression of transposons. Directly binds piRNAs, a class of 24 to 30 nucleotide RNAs that are generated by a Dicer-independent mechanism and are primarily derived from transposons and other repeated sequence elements. Associates with secondary piRNAs antisense and PIWIL2/MILI is required for such association. To purchase this antibody, use the following link: http://www.stjohnslabs.com/piwil4-antibody-p-66699?filter_name=STJ25009

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This gene encodes D-2hydroxyglutarate dehydrogenase, a mitochondrial enzyme belonging to the FAD-binding oxidoreductase/transferase type 4 family. This enzyme, which is most active in liver and kidney but also active in heart and brain, converts D-2-hydroxyglutarate to 2-ketoglutarate. Mutations in this gene are present in D-2-hydroxyglutaric aciduria, a rare recessive neurometabolic disorder causing developmental delay, epilepsy, hypotonia, and dysmorphic features. To purchase this antibody, use the following link: http://www.stjohnslabs.com/d2hgdh-antibody-p-68883?filter_name=STJ27209

Customer Review For CAMSAP3 Polyclonal Antibody (STJ46575)

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Microtubule minus-end binding protein that acts as a regulator of non-centrosomal microtubule dynamics and organization. Specifically required for the biogenesis and the maintenance of zonula adherens by anchoring the minus-end of microtubules to zonula adherens and by recruiting the kinesin KIFC3 to those junctional sites. May regulate the nucleation and the polymerization of microtubules. Indirectly, through the microtubule cytoskeleton, may regulate the organization of cellular organelles including the Golgi and the early endosomes. To purchase this antibody, use the following link: http://www.stjohnslabs.com/camsap3-antibody-n-term-p-78995?filter_name=STJ46575

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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

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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).[2][3] 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. Anti-α-tubulin -http://www.stjohnslabs.com/a-tubulin-antibody-p-98572 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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Antibody Customer Review For HIF-1α Polyclonal Antibody (STJ93498)

St John's Laboratory Ltd

Immunofluorescence Antibody Validation Report for Anti-α-tubulin Antibody (ST...

St John's Laboratory Ltd

Immunohistochemistry Antibody Validation Report for Anti-HIF-1α Antibody (STJ...

St John's Laboratory Ltd

Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB and EP300. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP. Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia. Anti-HIF-1α-http://www.stjohnslabs.com/hif-1a-antibody-p-92614 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunohistochemistry Antibody Validation Report for Anti-Claudin-5 Antibody (...

St John's Laboratory Ltd

Claudins are a family of proteins that are the most important components of the tight junctions, where they establish the paracellular barrier that controls the flow of molecules in the intercellular space between the cells of an epithelium. They have four transmembrane domains, with the N-terminus and the C-terminus in the cytoplasm. Anti-Claudin-5 -http://www.stjohnslabs.com/claudin-5-antibody-p-95192 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunofluorescence Antibody Validation Report for Anti-Claudin-5 Antibody (ST...

St John's Laboratory Ltd

Immunofluorescence Antibody Validation Report for Anti-GAPDH Antibody (STJ96931)

St John's Laboratory Ltd

Immunohistochemistry Antibody Validation Report for Anti-α-tubulin Antibody (...

St John's Laboratory Ltd

Immunocytochemistry Antibody Customer Review for Anti β-actin Antibody (STJ96...

St John's Laboratory Ltd

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. Beta actin is usually used as a loading control, for among others, the integrity of cells, protein degradation, in PCR and Western blotting. Its molecular weight is approximately 42 kDa. Anti β-actin - http://www.stjohnslabs.com/b-actin-antibody-p-98565?filter_name=STJ96930 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunofluorescence Antibody Validation Report for Anti-β-actin Antibody (STJ9...

St John's Laboratory Ltd

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[3][4] that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus. Beta actin is usually used as a loading control, for among others, the integrity of cells, protein degradation, in PCR and Western blotting. Its molecular weight is approximately 42 kDa. Anti-β-actin -http://www.stjohnslabs.com/b-actin-antibody-p-98565 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunohistochemistry Antibody Validation Report for Anti-GAPDH Antibody (STJ9...

St John's Laboratory Ltd

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. Anti-GAPDH-http://www.stjohnslabs.com/gapdh-antibody-p-98566 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunofluorescence Antibody Validation Report for Anti-CD81 Antibody (STJ96759)

St John's Laboratory Ltd

Immunohistochemistry Antibody Validation Report for Anti-Cleaved-PARP-1 (D214...

St John's Laboratory Ltd

Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. With EEF1A1 and TXK, forms a complex that acts as a T-helper 1 (Th1) cell-specific transcription factor and binds the promoter of IFN-gamma to directly regulate its transcription, and is thus involved importantly in Th1 cytokine production. Required for PARP9 and DTX3L recruitment to DNA damage sites. Anti-Cleaved-PARP-1 (D214)-http://www.stjohnslabs.com/cleaved-parp-1-d214-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunofluorescence Antibody Validation Report for Anti-Cleaved-PARP-1 (D214) ...

St John's Laboratory Ltd

Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. With EEF1A1 and TXK, forms a complex that acts as a T-helper 1 (Th1) cell-specific transcription factor and binds the promoter of IFN-gamma to directly regulate its transcription, and is thus involved importantly in Th1 cytokine production. Required for PARP9 and DTX3L recruitment to DNA damage sites. PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites. / NAD+ + (ADP-D-ribosyl)(n)-acceptor = nicotinamide + (ADP-D-ribosyl)(n+1)-acceptor. Anti-Cleaved-PARP-1 (D214) -http://www.stjohnslabs.com/cleaved-parp-1-d214-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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