RELA (phospho S529) polyclonal antibody
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic phosphopeptide of RELA.
Immunogen
Synthetic phosphopeptide (conjugated with KLH) corresponding to residues surrounding S529 of human RELA.
Sequence
PNGLLpSGDEDFC
Host
Rabbit
Reactivity
Human
Specificity
This phospho specific polyclonal antibody is specific to phosphorylated pS529 human p65. Reactivity with non-phosphorylated p65 is minimal.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:5000-1:10000)
Western Blot (1:1000)
The optimal working dilution should be determined by the end user.Storage Buffer
In antiserum (0.1% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Anti-pS529 shows phospho RELA staining in carcinoma cells.
Immunoblot of total protein lysates from various human head and neck tumors shows phospho RELA staining in tumor cell lines using p65.
Lanes 1 - 6 contain protein lysates from human squamal carcinoma cell lines.
Lane 7 is a protein lysate from a primary culture of human keratinocytes.
Lane 8 contains protein lysate from ATCC SCC9 cells (also a head and neck squamal carcinoma).
Lane 9 contains lysate from EGF-induced human derived A-431 cells.
Lane 10 (not shown) contains a molecular weight standard.
Concurrent staining with anti-beta microtubulin (not shown) was used to confirmequal protein loading in all lanes.
HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method.
Data contributed by Yu, M., NIH, personal communication.Western Blot (Cell lysate)
TNF Induces phosphorylation of RELA in KBM-5 cells.
Nuclear protein lysates prepared after 0, 5, 10, 15, 30 and 60 minutes of 0.1 nM TNF treatment of KBM-5 cells shows inducible phosphorylation using RELA (phospho S529) polyclonal antibody (Cat # PAB10337).
Anti-beta actin staining confirms loading of equivalent amounts of protein.
HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method.
Data contributed by Aggarwal BB, personal communication.Enzyme-linked Immunoabsorbent Assay
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Gene Info — RELA
Entrez GeneID
5970Gene Name
RELA
Gene Alias
MGC131774, NFKB3, p65
Gene Description
v-rel reticuloendotheliosis viral oncogene homolog A (avian)
Omim ID
164014Gene Ontology
HyperlinkGene Summary
NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA, or RELB (MIM 604758) to form the NFKB complex. The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NFKB complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).[supplied by OMIM
Other Designations
nuclear factor of kappa light polypeptide gene enhancer in B-cells 3|v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 (p65))|v-rel reticuloendotheliosis viral oncogene homolog
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Interactome
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Pathway
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Disease
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Publication Reference
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Piceatannol inhibits TNF-induced NF-kappaB activation and NF-kappaB-mediated gene expression through suppression of IkappaBalpha kinase and p65 phosphorylation.
Kazuhiro Ashikawa, Sekhar Majumdar, Sanjeev Banerjee, Alok C. Bharti, Shishir Shishodia, Bharat B. Aggarwal
The Journal of Immunology 2002 Dec; 169(11):6490.
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Control of oncogenesis and cancer therapy resistance by the transcription factor NF-kappaB.
Baldwin AS.
The Journal of Clinical Investigation 2001 Feb; 107(3):241.
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Series introduction: the transcription factor NF-kappaB and human disease.
Baldwin AS Jr.
J Clin Invest 2001 Jan; 107(1):3.
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Piceatannol inhibits TNF-induced NF-kappaB activation and NF-kappaB-mediated gene expression through suppression of IkappaBalpha kinase and p65 phosphorylation.
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