Product Pathways - NF-kappaB Signaling
IkappaB-alpha (112B2) Mouse mAb (Carboxy-terminal Antigen) #9247
| Applications | Reactivity | Sensitivity | MW (kDa) | Source | Isotype |
|---|---|---|---|---|---|
| W IP | H M R Mk | Endogenous | 39 | Mouse | IgG2a |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
IkappaB-alpha (112B2) Mouse mAb (Carboxy-terminal Antigen) detects endogenous levels of total IkappaB-alpha protein.
Source / Purification
Monoclonal antibody is produced by immunizing mice with a synthetic peptide (KLH-coupled) corresponding to residues near the carboxy-terminus of human IkappaB-alpha.
Western Blotting
Western blot analysis of extracts from Hela cells, treated with TNF-alpha (#2169, 10 ng/ml) for the indicated times, using Phospho-IkappaBalpha (Ser32) (14D4) Rabbit mAb #2859 (upper) or IkappaBalpha (112B2) Mouse mAb (Carboxy-terminal Antigen) (lower).
IP
Western blot analysis of extracts from HT29 cells (lane 1), protein A alone immunoprecipitation (lane 2), immunoprecipitation with nonspecific mouse IgG2a (lane 3), or immunoprecipitation with IkappaBalpha (112B2) Mouse mAb (Carboxy-terminal Antigen) (lane 4). Western blot was performed using IkappaBalpha Antibody #9242.
Background
The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκB-α at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκB-α phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8). Because phosphorylation of IκB-α at Ser32/36 is essential for release of active NF-κB, phosphorylation at this site is an excellent marker of NF-κB activation (1-3).
- Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
- Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
- Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
- Brown, K. et al. (1995) Science 267, 1485-8.
- Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.
Application References
- Gao, H. et al. (2004) Identification of beta-arrestin2 as a G protein-coupled receptor-stimulated regulator of NF-kappaB pathways. Mol. Cell 14, 303-317. This article references the use of IkappaB-alpha (112B2) Mouse mAb (Carboxy-terminal Antigen) in the following applications: IP Western Blotting
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Companion Products
- 4812 IκBα (44D4) Rabbit mAb
- 4814 IκB-α (L35A5) Mouse mAb (Amino-terminal Antigen)
- 9242 IkappaB-alpha Antibody
- 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
- 9246 Phospho-IκBα (Ser32/36) (5A5) Mouse mAb
- 4764 NF-κB p65 (C22B4) Rabbit mAb
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 9243 NF-κB Control Cell Extracts
- 9936 NF-κB Pathway Sampler Kit
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
- 7076 Anti-mouse 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
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
