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Product Pathways - NF-kB Signaling

NF-κB1 p105/p50 Antibody #3035

Item# Description List Price Web Price Qty
3035S NF-κB1 p105/p50 Antibody - 100 µl $383.00
$344.70
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Source
W Human, Monkey Endogenous 50 Active form. 120 Precursor. Rabbit
IP
ChIP

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP

Directions For Use

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

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 animals with a synthetic peptide corresponding to amino acids at the amino-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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30ng/ml) for 1 hour and either NF-κB1 p105/p50 Antibody #3035 or 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.


Background

Transcription factors of the nuclear factor κB (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).

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.

Application References

  • Kwong, C. et al. (2011) J Immunol 186, 1781-9. Applications: Western Blotting.
  • Rajendrasozhan, S. et al. (2010) Am J Physiol Lung Cell Mol Physiol 298, L197-209. Applications: Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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