Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

NF-κB Family Member Antibody Sampler Kit #4766

Kit Includes Quantity Applications Reactivity MW (kDa) Source
NF-κB p65 (C22B4) Rabbit mAb # 4764 40 microliters W IHC-P IF-IC F H M R Mk B (Dg) 65 Rabbit
NF-κB p65 Antibody # 3034 40 microliters W IP H M R Mk Mi Hm (Dg) 65 Rabbit
RelB (C1E4) Rabbit mAb # 4922 40 microliters W IP H M R Mk 70 Rabbit
c-Rel Antibody # 4727 40 microliters W IP IHC-P IF-IC F H Mk 78 Rabbit
NF-κB1 p105/p50 Antibody # 3035 40 microliters W IP H Mk 50 Active form. 120 Precursor. Rabbit
NF-κB1 p105 Antibody # 4717 40 microliters W IP H M R Mk Mi B 120 Rabbit
NF-κB2 p100/p52 Antibody # 4882 40 microliters W IP H M R Mk 52 active form. 120 precursor. Rabbit
NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) # 3017 40 microliters W IHC-P F H Mk 52 active form. 120 precursor. Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Mi=Mink  Hm=Hamster  B=Bovine  Dg=Dog

Specificity / Sensitivity

Each antibody in this kit recognizes endogenous levels of its target protein regardless of post-translational modification state such as phosphorylation or acetylation. The NF-κB1 p105/p50 Antibody #3035 detects both the precursor protein p105 and its cleavage product p50, while the NF-κB1 p105 Antibody #4717 only detects p105 and will not cross-react with p50. Both the NF-κB2 p100/p52 Antibody #4882 and the NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017 will cross-react with the precursor protein p100 and its cleavage product p52.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and COS cells using NF-kB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017.

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 #3034 (bottom).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with TNF-α (12 ng/ml) for the indicated amounts of time, using NF-kB p105/p50 Antibody #3035.


Western Blotting

Western Blotting

Western blot analysis of extracts from K562, Raji and 293 cells using c-Rel Antibody #4727.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper), NF-κB p65 (C22B4) Rabbit mAb #4764 (middle), and NF-κB1 p105 Antibody #4717 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using NF-kB2 p100/p52 Antibody #4882.


Western Blotting

Western Blotting

Western blot analysis of extracts from Raji, THP-1 and BaF3 cells using RelB (C1E4) Rabbit mAb #4922.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to amino acid residues surrounding Ser276 of human NF-κB p65, near the carboxy terminus of human c-Rel, near the amino terminus of human NF-κB1 p105/p50, at the carboxy terminus of human NF-κB1 p105 and near the amino terminus of human NF-κB2 p100/p52. Antibodies are purified by Protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to amino acid residues near the amino terminus of human NF-κB p65, surrounding Ser424 of human RelB and near the amino terminus of human NF-κB2 p100/p52.

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.

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