PathScan® Total IκBα Sandwich ELISA Kit #7360
Filter:
- ELISA
Supporting Data
| REACTIVITY | H M |
Application Key:
- ELISA-ELISA
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
Product Information
Product Description
CST's PathScan® Total IκBα Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total IκBα protein. An IκBα Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-IκBα proteins are captured by the coated antibody. Following extensive washing, an IκBα Rabbit Antibody is added to detect the captured IκBα protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total IκBα protein.
*Antibodies in kit are custom formulations specific to kit.
*Antibodies in kit are custom formulations specific to kit.
Protocol
Specificity / Sensitivity
CST's PathScan® Total IkappaB-alpha Sandwich ELISA Kit detects endogenous levels of total IκBα protein. Using PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit #7355, a significant induction of phospho-IκBα (Ser32) in HeLa cells treated with TNF-α can be detected. However, the level of total IκBα (phospho- and nonphospho-), detected by this Sandwich ELISA Kit #7360, remains unchanged (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Human, Mouse
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 IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα 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).
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- 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.
- Chen, J. et al. (2008) J Cell Mol Med , .
Product Citations: 1
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