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Primary Antibodies  >  Transcription Factor Antibodies

NFkB p65 phospho S536 Antibody

Rabbit Polyclonal


100 µg


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Rockland's affinity purified anti-p65 (RelA) pS536 antibody was used at 5.0  ?g/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate positive staining of human kidney distal tubules and collecting ducts. Tissue was formalin-fixed and paraffin embedded.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.  Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.
Western blot using Rockland's affinity purified anti-p65 (RelA) pS536 antibody shows detection of p65 phosphorylated at S536.  The control blot (left lane) contains 100 ng of purified p65-GST fusion protein.  The band is seen (right lane) when this protein is phosphorylated at S536 by IKKß. Personal Communication.  J. Brady, NCI, Bethesda, MD.
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Synonyms: NFKB, nfkb, NF-kB, NF-kappaB, NFkappaB

NFkB p65 phospho S536 Antibody Properties

Anti-NFKB p65 (Rel A) pS536 (RABBIT) Antibody - 600-401-265
Target Species
Known Cross Reactivity
ELISA : 1:1,000 - 1:6,000
Western Blot : 1:200 - 1:2,000
Immunohistochemistry: 5 µg/ml
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
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NFkB p65 phospho S536 Antibody Description

This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. NFkB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells.  It was subsequently found in non-B cells in an inactive cytoplasmic form consisting of NFkB bound to IkB.  NFkB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NFKB1) subunits.  Other identified subunits include p52 (NFKB2), cRel, and RelB.  The p65, cRel, and RelB subunits are responsible for transactivation.  The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkB subunit p65, similar to p50/p65 heterodimers.  Lowlevels of p52 and p50 homodimers can also exist in cells.  The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkB-a.  IkB-a binds to the p65 subunit, preventing nuclear localization, and DNA binding.  Activators mediate a rapid phosphorylation of IkB by IkB kinase (IKK), which results in subsequent ubiquitination and proteolytic degradation.  NFkB is then transported to the nucleus, where it activates transcription of target genes through binding to NFkB target sequences within the promoter.  The HTLV-I protein Tax can induce constitutive NFkB activation through phosphorylation of both I?B-a and I?B-ß.  The transforming protein Tax inhibits p53 transcriptional activity through the NFkB signaling pathway, specifically via the p65 (RelA) subunit.  The inhibition of p53 activity is dependent upon phosphorylation of p65 (RelA) at S536 by the upstream kinase IKKß.
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S536 of human p65 (RelA) protein.
Immunogen Type
Post Translational Modification
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and western blotting.  Specific conditions for reactivity should be optimized by the end user.  By western blot, a band approximately 65 kDa in size corresponding to phosphorylated p65 (RelA) protein is expected in the appropriate cell lysate or extract.  This phospho-specific polyclonal antibody reacts with human p65 (RelA) pS536 and shows minimal reactivity by western blot with non-phosphorylated p65 (RelA) and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide.
This product was affinity purified from monospecific antiserum by immunoaffinity chromatography using phospho-peptide coupled to agarose beads followed by solid phase adsorption against nonphospho-peptide. This antibody is specific for human p65 (RelA) protein phosphorylated at S536.  A BLAST analysis was used to suggest cross reactivity with p65 (RelA) from human, mouse and rat based on 100% homology with the immunizing sequence.  Cross reactivity with p65 (RelA) from other sources has not been determined.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Jeong S.J., Pise-Masison C.A., Radonovich M.F., Park H.U., and Brady J.N. (2005) A Novel NF-?B Pathway Involving IKKß? and p65/RelA Ser-536 Phosphorylation Results in p53 Inhibition in the Absence of NFkB Transcriptional Activity. J. Biol. Chem. 280 (11): 10326-10332. Baldwin, A.S. (2001) Control of oncogenesis and cancer therapy resistance by the transcription factor NFkB. J. Clinical Investigation 107: 241-246. Baldwin, A.S. (2001) Series introduction: the transcription factor NFkB and human disease. J. Clinical Investigation 107: 3-6.
Specific Reference
Manna SK, Aggarwal RS, Sethi G, et al. Morin (3,5,7,2’,4’-Pentahydroxyflavone) abolishes nuclear factor-kappaB activation induced by various carcinogens and inflammatory stimuli, leading to suppression of nuclear factor-kappaB-regulated gene expression and up-regulation of apoptosis. Clin. Cancer Res. 2007 Apr 1;13(7):2290-2297.
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