Product Pathways - NF-kB Signaling
Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033
PhosphoSitePlus® protein, site, and accession data: NFkB-p65
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | H M R Hm Mk Pg (Dg) | Endogenous | 65 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Hm=Hamster
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
Specificity / Sensitivity
Phospho-NF-kappaB p65 (Ser536) (93H1) Rabbit mAb detects NF-κB p65 only when phosphorylated at Ser536. It does not cross-react with the p50 subunit or other related proteins.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser536 of human NF-κB p65.
Western Blotting
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or TNF-α treated (#2169, 20 ng/ml for 5 minutes), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (upper) or NF-κB p65 Antibody #3034 (lower).
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 (upper) and NF-κB p65 (C22B4) Rabbit mAb #4764 (lower).
Flow Cytometry
Flow cytometric analysis of HeLa cells, untreated (blue) or TNF-α-treated (green), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb compared to a nonspecific negative control antibody (red).
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).
- Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
- Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
- Haskill, S. et al. (1991) Cell 65, 1281-9.
- Thompson, J.E. et al. (1995) Cell 80, 573-82.
- Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Senftleben, U. et al. (2001) Science 293, 1495-9.
- Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
- Xiao, G. et al. (2001) Mol Cell 7, 401-9.
Application References
- Lacasa, D. et al. (2007) Endocrinology 148, 868-77. Applications: Western Blotting
- Suzuki, S. et al. (2007) J Biol Chem 282, 25177-81. Applications: Western Blotting
- Lou, H. and Kaplowitz, N. (2007) J Biol Chem 282, 29470-81. Applications: Western Blotting
- Peiser, M. et al. (2008) J Leukoc Biol 83, 1118-27. Applications: ELISA
- Luedde, T. et al. (2008) Proc Natl Acad Sci U S A 105, 9733-8. Applications: Western Blotting
- Vince, J.E. et al. (2008) J Cell Biol 182, 171-84. Applications: Western Blotting
- Yadav, U.C. et al. (2009) Invest Ophthalmol Vis Sci 50, 2276-82. Applications: IF-P (paraffin)
- Dai, P. et al. (2009) J Immunol 182, 3450-60. Applications: Western Blotting
- Milsom, M.D. et al. (2009) Blood 113, 5111-20. Applications: Western Blotting
- Liu, M. et al. (2009) Am J Pathol 174, 1910-20. Applications: Western Blotting
- Yamanaka, Y. et al. (2009) Blood 114, 3265-75. Applications: Western Blotting
- Solt, L.A. et al. (2009) J Biol Chem 284, 27596-608. Applications: Western Blotting
- Wang, H. et al. (2009) J Immunol 183, 4755-63. Applications: IHC-F (floating)
- Xie, S. et al. (2010) J Immunol 184, 2289-96. Applications: Flow Cytometry IF-IC (In Cells) Western Blotting
- Kwong, C. et al. (2011) J Immunol 186, 1781-9. Applications: Western Blotting
- Stairs, D.B. et al. (2011) Cancer Cell 19, 470-83. Applications: Western Blotting
- Dental, C. et al. (2011) J Virol , . Applications: Flow Cytometry
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
- 3039 Phospho-NF-κB p65 (Ser468) Antibody
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 4812 IκBα (44D4) Rabbit mAb
- 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
- 4954 RelB Antibody
- 4727 c-Rel Antibody
- 3035 NF-κB1 p105/p50 Antibody
- 4806 Phospho-NF-κB p105 (Ser933) (18E6) Rabbit 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
- 8902 Human Tumor Necrosis Factor-α (hTNF-α)
- 8214 PhosphoPlus® NF-κB p65/RelA (Ser536) Antibody Duet
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.