NF-κB p65 (C22B4) Rabbit mAb

4764:

Flow , IHC / Paraffin , Immunofluorescence , Western Blotting

NF-κB p65 (C22B4) Rabbit mAb detects endogenous levels of total NF-κB p65 protein. This antibody may also cross-react with the p50 subunit.

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human NF-κB p65.

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) and NF-κB p65 (C22B4) Rabbit mAb (lower).

Western Blotting

Western blot analysis of extracts from HeLa (human), PC12 (rat) and Neuro2A (mouse) cell lines using NF-κB p65 (C22B4) Rabbit mAb.

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence^® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence^® NF-κB p65 siRNA II (+), or SignalSilence® NF-κB p65 siRNA I #6261 (+), using NF-κB p65 (C22B4) Rabbit mAb #4764 and α-Tubulin (11H10) Rabbit mAb #2125. NF-κB p65 (C22B4) Rabbit mAb confirms silencing of NF-κB p65 and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of NF-κB p65 siRNA.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human melanoma using NF-κB p65 (C22B4) Rabbit mAb.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cells, untreated (left) or treated with TNF-α (right), using NF-κB p65 (C22B4) Rabbit mAb.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using NF-κB p65 (C22B4) RmAb #4764 in the presence of control peptide (left) or NF-κB p65 Blocking Peptide #1061 (right).

Flow Cytometry

Flow cytometric analysis of OVCAR8 cells using NF-κB p65 (C22B4) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or TNF-α-treated (#8902, 20ng/ml for 20 min, right), using NF-κB p65 (C22B4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor^® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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).

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.

• Hasler, U. et al. (2008) J Biol Chem 283, 28095-105. Applications: Western Blotting
• Hideshima, T. et al. (2009) Blood 113, 5228-36. Applications: IHC-P (paraffin) Western Blotting
• Xie, S. et al. (2010) J Immunol 184, 2289-96. Applications: Western Blotting
• Ikner, A. and Ashkenazi, A. (2011) J Biol Chem 286, 21546-54. Applications: Western Blotting
• Stairs, D.B. et al. (2011) Cancer Cell 19, 470-83. Applications: IC-IF 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 !

• 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
• 3039 Phospho-NF-κB p65 (Ser468) Antibody
• 3034 NF-κB p65 Antibody
• 4954 RelB Antibody
• 4727 c-Rel Antibody
• 3035 NF-κB1 p105/p50 Antibody
• 4882 NF-κB2 p100/p52 Antibody
• 4812 IκBα (44D4) Rabbit mAb
• 4814 IκBα (L35A5) Mouse mAb (Amino-terminal Antigen)
• 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
• 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
• 1061 NF-κB p65 Blocking Peptide
• 9243 NF-κB Control Cell Extracts
• 9936 NF-κB Pathway Sampler Kit
• 8112 SignalStain^® Antibody Diluent
• 8114 SignalStain^® Boost IHC Detection Reagent (HRP, Rabbit)
• 5425 Normal Goat Serum
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 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)

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