Product Pathways - NF-kB Signaling

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Phospho-NF-κB p65 (Ser536) Antibody #3031

Applications	Reactivity	Sensitivity	MW (kDa)	Source
W	H M R Mk (Dg) (Pg)	Endogenous	65	Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dg=Dog  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

3031:

Western Blotting

Specificity / Sensitivity

Phospho-NF-kappaB p65 (Ser536) Antibody detects NF-kappaB p65 only when phosphorylated at serine 536. The antibody does not cross-react with the p50 subunit or other related proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser536 of human NF-kappaB p65. Antibodies are purified by protein A and/or peptide affinity chromatography.

Background

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 translocates 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

• Haller, D. et al. (2002) J Biol Chem 277, 38168-78. Applications: Western Blotting
• Strassheim, D. et al. (2004) J Immunol 172, 5727-33. Applications: Western Blotting
• Haller, D. et al. (2003) J Biol Chem 278, 23851-60. Applications: IHC-P (paraffin) Western Blotting
• Ruiz, P.A. et al. (2004) J Biol Chem 279, 36103-11. Applications: Western Blotting
• Müller, I. et al. (2003) Cancer Res 63, 1772-5. Applications: Western Blotting
• Anwar, K.N. et al. (2004) J Immunol 173, 6965-72. 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!

Companion Products

• 4764 NF-κB p65 (C22B4) Rabbit mAb
• 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
• 4954 RelB Antibody
• 4727 c-Rel Antibody
• 3035 NF-κB1 p105/p50 Antibody
• 4882 NF-κB2 p100/p52 Antibody
• 9243 NF-κB Control Cell Extracts
• 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

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