Cell Signaling Technology

Product Pathways - NF-kB Signaling

Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Biotinylated) #4025

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M R Hm Mk Pg (Dg) Endogenous 65 Rabbit IgG

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

Protocols

Specificity / Sensitivity

Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Biotinylated) detects NF-κB p65 protein only when phosphorylated at Ser536. It does not cross-react with the p50 subunit or other related proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser536 of human NF-κB p65 protein.

Western Blotting

Western Blotting

Western blot analysis of NF-κB Control Cell Extracts #9243 (HeLa untreated or treated with hTNF-α #8902, 20 ng/ml, 5 min.), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Biotinylated).

Description

This Cell Signaling Technology (CST) antibody is conjugated to biotin under optimal conditions. The unconjugated Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 reacts with human, mouse, rat, monkey, hamster, and pig phospho-NF-κB. CST expects that Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Biotinylated) will also recognize phospho-NF-κB in these species.

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 translocates 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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For Research Use Only. Not For Use In Diagnostic Procedures.

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