RelA/NFkB p65 [p Ser276] Antibody - BSA Free

Western Blot: RelA/NFkB p65 [p Ser276] Antibody [NBP1-77827] - Probed against Normal (Lane 1) and TNFalpha (Lane 2) Stimulated HeLa whole cell lysates. A band was observed between 55 and 72 kD corresponding to the expected MW of NFkB p65. The observed higher MW bands have not been characterized.

• Reactivity: Hu
• Applications: WB, ELISA, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

RelA/NFkB p65 [p Ser276] Antibody - BSA Free Summary

• Description: This antibody RelA/NFkB p65 p Ser276 Antibody was prepared from monospecific antiserum by delipidation and defibrination
  
  Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: RelA/NFkB p65 peptide corresponding to an internal region near phospho Serine 276 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH). (Uniprot: Q04206)
• Modification: p Ser276
• Specificity: The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. This phospho specific polyclonal antibody is specific for phosphorylated pS276 human RelA/NFkB p65. Reactivity with non-phosphorylated pS267 is minimal. Cross reactivity with pS276 phosphorylated RelA/NFkB p65 from mouse, rat or other species has not been determined.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: RELA
• Purity: Whole Antiserum

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000-1:25000
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot 1:1000
• Application Notes: This product reacts with human pS276 p65 and shows minimal reactivity when tested by western blot with non-phosphorylated p65 and minimal reactivity when tested by ELISA against the non-phosphorylated form of the immunizing peptide. For immunoblotting a 1:1,000 dilution is recommended. A 65 kDa band corresponding to human p65 is detected. HeLa cells or TNF inducible KBM-5 cells can be used as a positive control. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.
• Control Peptide:

  RelA/NFkB p65 Protein (NBP1-97002)
  RelA/NFkB p65 Protein (NBP1-97003)

Reactivity Notes

This phospho specific polyclonal antibody is specific for phosphorylated pS276 human RelA/NFkB p65. Reactivity with non-phosphorylated pS267 is minimal. Cross reactivity with pS276 phosphorylated RelA/NFkB p65 from mouse, rat or other species has not been determined.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Whole Antiserum

Alternate Names for RelA/NFkB p65 [p Ser276] Antibody - BSA Free

• MGC131774
• NF kB p65 phospho
• nf kb p65
• NFkB p65 phospho
• NF-kB p65 phospho
• NFkB p65
• NF-kB p65
• NFKB3
• NFKB3v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor ofkappa light polypeptide gene enhancer in B-cells 3 (p65))
• Nuclear factor NF-kappa-B p65 subunit
• Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3transcription factor p65
• p65
• p65RelA
• phospho NF kB p65
• phospho NFkB p65
• phospho NF-kB p65
• rela p65
• RelA
• v-rel reticuloendotheliosis viral oncogene homolog A (avian)
• v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight polypeptide gene enhancer in B-cells 3, p65

Background

NF'B 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 NF'B bound to I'B. NF'B 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 NF'B subunit p65, similar to p50/p65 heterodimers. Low levels 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 I'B-a. I'B-a 'binds to the p65 subunit preventing nuclear localization and DNA binding. Activators mediate a rapid phosphorylation of I'B by I'B kinase (IKK) which results in subsequent ubiquitination and proteolytic degradation. NF'B is then transported to the nucleus, where it activates transcription of target genes through binding to NF'B target sequences within the promoter. The HTLV-I protein Tax can induce constitutive NF'B activation through phosphorylation of both I'B-a and I'B-B. The transforming protein Tax inhibits p53 transcriptional activity through the NF'B 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 IKKB.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for RelA/NFkB p65 Antibody (NBP1-77827)(2)

Publications using NBP1-77827

• Sheldon M, Rice DS, D'Arcangelo G et al. Toll-like receptor 2 mediates apolipoprotein CIII-induced monocyte activation. Circ Res 2008-12-01 [PMID: 18974386]
• Ashikawa K, Majumdar S, Banerjee S et al. Piceatannol inhibits TNF-induced NF-kappaB activation and NF-kappaB-mediated gene expression through suppression of IkappaBalpha kinase and p65 phosphorylation. J Immunol. 2002-12-01 [PMID: 12444159]

Bioinformatics

• Gene Symbol: RELA
• Uniprot:
  • Q04206()
  • Q04206()
  • Q04206()
  • Q04206()
  • Q99496(Human)