NF-κB1 p105/p50 Antibody

3035:

ChIP Agarose , ChIP Magnetic , Immunoprecipitation , Western Blotting

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

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids at the amino-terminus of human NF-kappaB p105.

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 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.

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30ng/ml) for 1 hour and either 20 μl of NF-κB1 p105/p50 Antibody #3035 or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP^® Human IkB-alpha Promoter Primers #5552, human IAP2 promoter primers, and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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).

Following IKK-mediated phosphorylation of p105 NF-κB at multiple sites (Ser921, 923, 927, and 932) on its carboxy-terminus, SCF/β-TrCP mediated processing produces the 50 kDa 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.

• Kwong, C. et al. (2011) J Immunol 186, 1781-9. Applications: Western Blotting

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• 4806 Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb
• 4808 Phospho-NF-κB p105 (Ser933) (178F3) Rabbit mAb (IHC Specific)
• 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
• 3036 Phospho-NF-κB p65 (Ser536) (7F1) Mouse mAb
• 3039 Phospho-NF-κB p65 (Ser468) Antibody
• 4812 IκBα (44D4) Rabbit mAb
• 4954 RelB Antibody
• 4727 c-Rel Antibody
• 2682 IKKα Antibody
• 2370 IKKβ (2C8) Rabbit mAb
• 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
• 2695 IKKγ (DA10-12) Mouse mAb
• 4882 NF-κB2 p100/p52 Antibody
• 4810 Phospho-NF-κB2 p100 (Ser866/870) Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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