Cell Signaling Technology

Product Pathways - NF-kB Signaling

NF-κB p65 Antibody #3034

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Hm Mk Mi (Dg) Endogenous 65 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  Dg=Dog
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

NF-κB p65 Antibody detects endogenous levels of total NF-κB p65. This antibody may also cross-react with the p50 subunit.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids surrounding Ser276 of human NF-κB p65.

Western Blotting

Western Blotting

Western blot anlaysis of extracts from HeLa cells, untreated or TNF-α-treated (#2169, 20 ng/ml) for the indicated times, using Phospho-NF-κB p65 (Ser536) Antibody #3031 (top) or NF-κB p65 Antibody (bottom).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with nontargeted (-) or targeted (+) siRNA. p65 was detect using NF-κB p65 Antibody #3034, and p42 was detected using p42 MAPK Antibody #9108. The NF-κB p65 Antibody confirms silencing of p65 expression, and the p42 MAPK Antibody is used to control for loading and siRNA specificity.

Background

Transcription factors of the nuclear factor κ B (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 the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which is then translocated to the nucleus (9-11).

  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.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

* Offer is for U.S. customers only. There is a limit of one (1) free trial size of product 8242 per order, subject to availability. Offer is good on orders placed December 12, 2011 through March 12, 2012.

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