Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

Phospho-NF-κB p65 (Ser276) Antibody #3037

Applications Reactivity MW (kDa) Source
W IHC-P H M R (B) (Dg) 65-80 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-NF-κB p65 (Ser276) Antibody detects NF-κB p65 only when phosphorylated at Ser276. It may also cross-react with c-Rel and NF-κB1 p105/p50 when phosphorylated at the homologous residues (Ser267 and Ser337, respectively).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser276 of human NF-κB p65. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with calyculin A #9902 (50 nM) for 10 minutes, using Phospho-NF-κB p65 (Ser276) Antibody (upper) or NF-κB p65 Antibody #3034 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-NF-κB p65 (Ser 276) Antibody in the presence of control peptide (left) or Phospho-NF-κB p65 (Ser276) Blocking Peptide #1437 (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or Calf Intestinal Phosphatase (CIP) treated (right), using Phospho-NF-kappaB p65 (Ser276) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-NF-κB p65 (Ser276) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysisof paraffin-embedded human colon carcinoma using Phospho-NF-kappaB p65 (Ser276) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysisof paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-NF-κB p65 (Ser276) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded PC-3 cells using Phospho-NF-κB p65 (Ser276) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-NF-κB p65 (Ser276) Antibody in the presence of control peptide (left) or Phospho-NF-κB p65 (Ser276) Blocking Peptide #1437 (right).

Background

Transcription factors of the nuclear factor kappaB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50) and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through an ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKK-α (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which is then translocated to the nucleus (9-11).

Phosphorylation of NF-κB p65 by PKA and/or MSK1 at Ser276 allows for increased interaction with the transcriptional coactivator p300/CBP to further enhance the transcriptional activity of this transcription factor (12-14)

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.
  12. Zhong, H. et al. (1997) Cell 89, 413-424.
  13. Zhong, H. et al. (1998) Mol. Cell 1, 661-671.
  14. Vermeulen, L. et al. (2003) EMBO J. 22, 1313-1324.

Application References

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