Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

Phospho-NF-κB p65 (Ser536) (7F1) Mouse mAb #3036

Applications Reactivity Sensitivity MW (kDa) Source Isotype
W H M R Mk Mi (Dg) Endogenous 65 Mouse IgG2b

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Mi=Mink  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-kappaB p65 (Ser536) (7F1) Mouse mAb detects NF-kappaB p65 only when phosphorylated at serine 536. It does not cross-react with the p50 subunit or other related proteins.

Source / Purification

Monoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser536 of human NF-kappaB p65.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or TNF-alpha-treated (#2169, 20ng/ml), using Phospho-NF-kappaB p65 (Ser536) (7F1) Mouse mAb (upper) or NF-kappaB p65 Antibody #3034 (lower).

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).

  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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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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