Customer Review For NFκB-p65 Polyclonal Antibody (STJ94468)

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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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HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: NFκB-p65 Polyclonal Antibody (STJ94468)

WHAT WAS USED?
Primary antibody: STJ94468. NFκB-p65 Polyclonal
Antibody. 100µl
Provider: St John's Laboratory
Dilution ratio: 1:1000
Application: Western Blot.
Image: 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.

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 STJ94468.
NFκB-p65
Polyclonal
Antibody
1:1000
1:10000 65KD 30S
2 A549 lysate 10 µg 1:10000 65KD 30S

WHAT DID THE CUSTOMER THINK?
Testing results were provided by BPI laboratory. They
stated that “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”.

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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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The Survivin protein also known as BIRC5 (baculoviral inhibitor of apoptosis repeat-containing 5) is a member of the inhibitor of apoptosis family. It is encoded by the BIRC5 gene in humans. It is a multitasking protein through both promoting cell proliferation and preventing cell death. It is thought to regulate apoptosis through inhibiting caspase activation. The Survivin monoclonal antibody detects endogenous Survivin protein.

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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. Anti-Phospho-NFκB-p65 (S536) -http://www.stjohnslabs.com/phospho-nfkb-p65-s536-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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Antibody Customer Review for Anti-Survivin Monoclonal Antibody (STJ97436)

St John's Laboratory Ltd

Immunofluorescence Antibody Validation Report for Anti-Phospho-NFκB-p65 (S536...

St John's Laboratory Ltd

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. Anti-Phospho-NFκB-p65 (S536) -http://www.stjohnslabs.com/phospho-nfkb-p65-s536-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Immunohistochemistry Antibody Validation Report for Anti-Phospho-NFκB-p65 (S5...

St John's Laboratory Ltd

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. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. Anti-Phospho-NFκB-p65 (S536)-http://www.stjohnslabs.com/phospho-nfkb-p65-s536-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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

St John's Laboratory Ltd

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

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

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-IL-6 Antibody (STJ96...

St John's Laboratory Ltd

Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. Acts on B-cells, T-cells, hepatocytes, hematopoietic progenitor cells and cells of the CNS. Required for the generation of T(H)17 cells. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation. Anti-IL-6-http://www.stjohnslabs.com/il-6-antibody Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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

St John's Laboratory Ltd

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

Immunohistochemistry Antibody Validation Report for Anti-β-actin Antibody (ST...

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

Western Blot Antibody Customer Review for Anti-Peroxin 2 Antibody (STJ95040)

St John's Laboratory Ltd

PEX2 is gene on chromosome 8q21.1 that encodes a peroxin, an integral membrane protein involved in peroxisome biosynthesis and integrity, which assembles membrane vesicles before translocation of matrix proteins. Defects of PEX2 cause peroxisome biogenesis disorder complementation group 5, Zelleger syndrome and a form of Refsum disease. Anti-Peroxin 2 - http://www.stjohnslabs.com/peroxin-2-antibody?filter_name=STJ95040 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

Immunofluorescence Antibody Validation Report for Anti-Histone H2B Antibody (...

St John's Laboratory Ltd

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Anti-Histone H2B -http://www.stjohnslabs.com/histone-h2b- antibody-p-98699 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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

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