Product Pathways - NF-kappaB Signaling
NF-κB Family Member Antibody Sampler Kit #4766
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| NF-κB p65 (C22B4) Rabbit mAb # 4764 | 40 microliters | W IHC-P IF-IC F | H M R Mk B (Dg) | 65 | Rabbit |
| NF-κB p65 Antibody # 3034 | 40 microliters | W IP | H M R Mk Mi Hm (Dg) | 65 | Rabbit |
| RelB (C1E4) Rabbit mAb # 4922 | 40 microliters | W IP | H M R Mk | 70 | Rabbit |
| c-Rel Antibody # 4727 | 40 microliters | W IP IHC-P IF-IC F | H Mk | 78 | Rabbit |
| NF-κB1 p105/p50 Antibody # 3035 | 40 microliters | W IP | H Mk | 50 Active form. 120 Precursor. | Rabbit |
| NF-κB1 p105 Antibody # 4717 | 40 microliters | W IP | H M R Mk Mi B | 120 | Rabbit |
| NF-κB2 p100/p52 Antibody # 4882 | 40 microliters | W IP | H M R Mk | 52 active form. 120 precursor. | Rabbit |
| NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) # 3017 | 40 microliters | W IHC-P F | H Mk | 52 active form. 120 precursor. | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 100 microliters | Goat |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Mi=Mink
Hm=Hamster
B=Bovine
Dg=Dog
Specificity / Sensitivity
Each antibody in this kit recognizes endogenous levels of its target protein regardless of post-translational modification state such as phosphorylation or acetylation. The NF-κB1 p105/p50 Antibody #3035 detects both the precursor protein p105 and its cleavage product p50, while the NF-κB1 p105 Antibody #4717 only detects p105 and will not cross-react with p50. Both the NF-κB2 p100/p52 Antibody #4882 and the NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017 will cross-react with the precursor protein p100 and its cleavage product p52.
Western Blotting
Western blot analysis of extracts from HeLa and COS cells using NF-kB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017.
Western Blotting
Western blot anlaysis of extracts from HeLa cells, untreated or TNF-α-treated (#2169, 20 ng/ml) for the indicated times, using Phospho-NF-κB p65 (Ser536) Antibody #3031 (top) or NF-κB p65 Antibody #3034 (bottom).
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or treated with TNF-α (12 ng/ml) for the indicated amounts of time, using NF-kB p105/p50 Antibody #3035.
Western Blotting
Western blot analysis of extracts from K562, Raji and 293 cells using c-Rel Antibody #4727.
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), NF-κB p65 (C22B4) Rabbit mAb #4764 (middle), and NF-κB1 p105 Antibody #4717 (lower).
Western Blotting
Western blot analysis of extracts from various cell types using NF-kB2 p100/p52 Antibody #4882.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to amino acid residues surrounding Ser276 of human NF-κB p65, near the carboxy terminus of human c-Rel, near the amino terminus of human NF-κB1 p105/p50, at the carboxy terminus of human NF-κB1 p105 and near the amino terminus of human NF-κB2 p100/p52. Antibodies are purified by Protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to amino acid residues near the amino terminus of human NF-κB p65, surrounding Ser424 of human RelB and near the amino terminus of human NF-κB2 p100/p52.
Background
Transcription factors of the nuclear factor kappaB (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 an ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKK-α (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which is then translocated 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
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Companion Products
- 9936 NF-κB Pathway Sampler Kit
- 3037 Phospho-NF-κB p65 (Ser276) Antibody
- 3039 Phospho-NF-κB p65 (Ser468) Antibody
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 4806 Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb
- 4808 Phospho-NF-κB p105 (Ser933) (178F3) Rabbit mAb (IHC Specific)
- 4810 Phospho-NF-κB2 p100 (Ser866/870) Antibody
- 9243 NF-κB Control Cell Extracts
- 2169 Tumor Necrosis Factor-α
- 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