Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Antibody Pair #7343

Kit Includes Volume Cap Color
IkB-alpha Mouse Capture Antibody (100X) 0.4 milliliters Pink
Phospho-IkB-alpha (Ser32) 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® Phospho-IκB-α (Ser32) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit #7355. 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 a Phospho-IκBα (Ser32) 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 Phospho-IκBα (Ser32) protein.*Antibodies in this kit are custom formulations specific to the kit.

Sandwich ELISA

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® Phopho-IκBα (Ser32) Sandwich ELISA Antibody Pair #7343 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).

  1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
  2. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
  3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
  4. Brown, K. et al. (1995) Science 267, 1485-8.
  5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  8. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.

Application References

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Companion Products

Rabbit Monoclonals are produced under license (granting certain rights, including those under U.S. Patent No. 5,675,063) from Epitomics, Inc.

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.

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