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NFkB p65 (RelA) Phospho S276 Antibody

Catalogue number:
100-401-264
Size:
100 µl
Product is available in:
  • UK
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POA
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Applications:

ELISA, Immunohistochemistry, Western Blot

Antibody Host:

Rabbit

Species Reactivity:

H. sapiens (Human)

Antibody Isotype:

Antiserum

Antibody Type:

Polyclonal

Immunogen:

NFkB p65 (Rel A) peptide corresponding to an internal region near phospho Serine 276 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH

Alternative Names:

rabbit anti-NFkB p65 pS276 Antibody, rabbit anti-RelA pS276 Antibody, NFKB, nfkb, NF-kB, NF-kappaB, NFkappaB, Anti-NF-kB antibody

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Product Description:

NF-kB 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-kB bound to IkB. NF-kB 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-kB 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 IkB-alpha. IkB-alpha 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. NF-kB is then transported to the nucleus, where it activates transcription of target genes through binding to NF-kB target sequences within the promoter. The HTLV-I protein Tax can induce constitutive NF-kB activation through phosphorylation of both IkB-alpha and IkB-beta. 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 IKKβ. Anti-NFkB antibody is ideal for Cell Biology, Nuclear Signaling, Neuroscience and Signal Transduction Research. Phospho NFkB antibody reacts human pS276 p65 and shows minimal reactivity by western blot with non-phosphorylated p65 and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. A 1:500 dilution has been used for staining p65 in human kidney tissue by IHC. Tissue was formalin fixed and paraffin embedded. Although not tested, this antibody is likely functional in immunoprecipitation. All conditions must be user optimized.

Rockland's anti-p65 pS276 antibody was d
Rockland's anti-p65 pS276 antibody was diluted 1:500 to detect p65 in human kidney tissue. Tissue was formalin fixed and paraffin embedded. No pre-treatment of sample was required. The image shows the localization of antibody as the precipitated red signal, with a hematoxylin purple nuclear counter stain.
 Anti-pS276 shows phospho p65 staining i
Anti-pS276 shows phospho p65 staining in carcinoma cells. Western blot of total protein lysates from various human head and neck tumors shows phospho p65 staining in tumor cell lines using phospho specific polyclonal anti-human pS276 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 and does not show significant levels of phosphorylated p65. 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 A431 cells. Lane 10 (not shown) contains a molecular weight standard. Concurrent staining with anti-beta microtubulin (not shown) was used to confirm equal protein loading in all lanes. HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method. Other detection methods will yield similar results. Data contributed by Yu, M., NIH, personal communication.