Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

NF-κB1 p105/p50 Antibody #3035

Applications Reactivity MW (kDa) Source
W IP H Mk 50 Active form. 120 Precursor. Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

NF-kappaB p105/p50 Antibody detects endogenous levels of the precurser protein p105 and its cleavage product p50.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to amino acids at the animo-terminus of human NF-kappaB p105.

Western Blotting

Western Blotting

Western blot analysis of extracts from Vero cells, untreated or treated with TNF-α #2169 (20 ng/ml) for the times indicated, using Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb #4806 (upper) and NF-κB p105/p50 Antibody #3035 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Hela cells, untreated or treated with TNF-alpha (12ng/ml) for the indicated amounts of time, using NF-kappaB p105/p50 Antibody.

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).

Following IKK-mediated phosphorylation of p105 NF-kappaB at multiple sites (Ser921, 923, 927, and 932) on its carboxy-terminus, SCFbeta-TrCP mediated processing produces the 50kDa active form p50 (12,13).

  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. Heissmeyer, V. et al. (2001) Mol Cell Biol 21, 1024-1035.
  13. Orian, A. et al. (2000) EMBO J 19, 2580-2591.

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