IKKa/b, phosphorylated (Ser176/180) Antibody Sampler Kit, BioAssay(TM)

Intended Use:\nThe Phospho-IKKa/b (Ser176/180) Antibody Sampler Kit contains reagents to examine protein levels of IKKa when phosphorylated at Ser176/180 and IKKb when phosphorylated at Ser177/181. The kit contains primary and secondary antibodies to perform four Western Blots with each antibody.\n\nBackground: \nThe NF-kB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IkB proteins (1-3). Most agents that activate NF-kB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degrada- tion of IkB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IkB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKa and IKKb serve as the catalytic subunits of the kinase. IKKg serves as the regulatory subunit (8-9). Activation of IKK depends on phosphorylation; Ser177 and Ser181 in the activation loop of IKKb (176 and 180 in IKKa) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).\n\nKit Components:\n1. Phospho-IKKa (Ser176)/IKKb (Ser177) (C84E11) Mab rabbit x human, 1x40ul\n2. Phospho-IKKa/b (Ser176/180) (16A6) Mab rabbit x human, 1x40ul\n3. Phospho-IKKa/b (Ser176/180) Pab rabbit x human, 1x40ul\n4. IgG (HRP) Pab Goat x Rabbit, 1x100ul\n\nApplications: \nSuitable for use in Western Blot. Phospho-IKKa/b (Ser176/180) (16A6) Mab rabbit x human has been tested in Immunohistochemistry (frozen and paraffin). Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000\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. \n

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SPECIFICATIONS

Catalog Number

I3000-11B

Size

1Kit

Applications

WB

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM NaCl, 100ug/ml BSA and 50% glycerol.

References

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. Zandi, E. et al. (1997) Cell 91, 243-52. 9. Karin, M. (1999) Oncogene 18, 6867-74. 10. DiDonato, J.A. et al. (1997) Nature 388, 548-54. 11. Mercurio, F. et al. (1997) Science 278, 860-6. 12. Johnson, L.N. et al. (1996) Cell 85, 149-58. 13. Delhase, M. et al. (1999) Science 284, 309-13.

Additional Info

Recognizes IKKa at 85kD only when phosphorylated at Ser176/180 and IKKb at 87kD only when phosphorylated at Ser177/181.\n1. Phospho-IKKa (Ser176)/IKKb (Ser177) (C84E11) Mab rabbit x human: recognizes human, mouse. Sequence homology: rat, monkey, bovine.\n2. Phospho-IKKa/b (Ser176/180) (16A6) Mab rabbit x human: recognizes human, mouse, rat, monkey. Sequence homlogy: bovine.\n3. Phospho-IKKa/b (Ser176/180) Pab rabbit x human recognzies human, mouse, rat, monkey.\n