IkB alpha, phosphorylated (Ser32/36)

The NF-kappaB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IkB proteins (1-3). Activation occurs via phosphorylation of IkB-alpha at Ser32 and Ser36, resulting in the release and nuclear translocation of active NF-kappaB (3-7). IkappB-alpha phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals, including inflammatory cytokines, growth factors and chemokines. Phosphorylation of IkappB-alpha at Ser32 and Ser36 has been shown to stimulate conjugation with ubiquitin followed by proteasome-mediated degradation of IkB, resulting in the release of active NF-kappaB. Kinases that phosphorylate IkB at these activating sites have been identified (8). Because phosphorylation of IkB-alpha at Ser32 is essential for release of active NF-kappaB, phosphorylation at this site is an excellent marker of NF-kappaB activation (1-3).\n\nApplications:\nSuitable for use in Western Blot. Other applications have not been tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000. Incubate membrane with diluted I2500-06A in TBS, 5% nonfat dry milk, 0.1% Tween-20 overnight at 4 degrees C with gentle shaking.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

I2500-06A

Size

100ul

Applications

WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, <0.02% sodium azide, 50% glycerol.

P Type

Mab

Purity

Ascites

Isotype

IgG1

References

1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-546. 2. Beg, A.A. et al. (1993) Genes Dev. 7, 2064-2070. 3. Finco, T.S. et al. (1994) PNAS USA 91, 11884-11888. 4. Brown, K. et al. (1995) Science 267, 1485-1488. 5. Brockman, J.A. et al. (1995) Mol. Cell. Biol. 15, 2809-2818. 6. Traenckner, E.B. et al. (1995) EMBO J. 14, 2876-2883. 7. Chen, Z.J. et al. (1996) Cell 84, 853-862. 8. Karin, M., Ben-Neriah, Y. (2000) Annu. Rev. Immunol. 18, 621-663. 9. Howe, C.J. et al. (2002) J. Biol. Chem. 277, 30469-30476. 10. Howe, C.L., et al.: Precipitation of the CaM-kinase in hydrogen peroxide-induced Ikb phosphorylation in human T lymphocytes. J. Biol. Chem. 277: 30469-30476 (2002).

Additional Info

Recognizes endogenous levels of human IkappaB-alpha only when phosphorylated at Ser32/36 at 40kD. Species Crossreactivity: mouse, rat and monkey.