Product Pathways - PathScan ELISA
PathScan® Total IκBα Sandwich ELISA Antibody Pair #7831
| Kit Includes | Volume | Cap Color |
|---|---|---|
| IkB-alpha Mouse Capture Antibody (100X) | 0.4 milliliters | Pink |
| IkB-alpha Rabbit Detection Antibody (100X) | 0.4 milliliters | Blue |
| Anti-rabbit IgG HRP-Linked Antibody (1000X) | 0.04 milliliters | Red |
Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.
Species Cross-Reactivity
H M
Reactivity Key: H=Human M=Mouse
Description
CST's PathScan® Total IκBα Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan®Total IκBα Sandwich ELISA Kit #7360. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The IκBα Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by an IκBα Detection Antibody and anti-Rabbit IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of total IκBα protein.*Antibodies in this kit are custom formulations specific to the kit.
Sandwich ELISA
The relationship between lysate protein concentration from untreated and TNF-α and IL-1β treated HeLa cells and the absorbance at 450 nm using PathScan® Total IκBα Sandwich ELISA Antibody Pair #7831 is shown. HeLa cells were treated with TNF-α and IL-1β for 5 minutes at 37°C and then lysed.
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).
- 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
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
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7360 PathScan® Total IκBα Sandwich ELISA Kit
- 7355 PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit
- 7343 PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Antibody Pair
- 4812 IκBα (44D4) Rabbit mAb
- 4814 IκBα (L35A5) Mouse mAb (Amino-terminal Antigen)
- 9247 IκBα (112B2) Mouse mAb (Carboxy-terminal Antigen)
- 9242 IκBα Antibody
- 9803 Cell Lysis Buffer (10X)
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.
