Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

Phospho-IKKα (Ser180)/IKKβ (Ser181) Antibody #2681

Applications Reactivity MW (kDa) Source
W H M (R) (B) 85 (IKKalpha). 87 (IKKbeta). Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-IKKalpha (Ser180)/IKKbeta (Ser181) Antibody detects IKKalpha and IKKbeta only when phosphorylated at serine 180 or serine181, respectively.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser180 of human IKK alpha. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated with TNF-alpha (#2169, 20 ng/ml) for the indicated times, using Phospho-IKKalpha (Ser180)/IKKbeta (Ser181) Antibody (upper) and control IKKalpha/beta antibodies (lower).

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase. IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends on phosphorylation; Ser177 and Ser181 in the activation loop of IKKβ (176 and 180 in IKKα) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).

  1. Baeuerle, P.A. et al. (1988) Science 242, 540-546.
  2. Beg, A.A. et al. (1993) Genes Dev. 7, 2064-2070.
  3. Finco, T.S. et al. (1994) Proc. Natl. Acad. Sci. 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. Zandi, E. et al. (1997) Cell 91, 243-252.
  9. Karin, M. et al. (1999) Oncogene 18, 6867-6874.
  10. DiDonato, J.A. et al. (1997) Nature 388, 548-554.
  11. Mercurio, F. et al. (1997) Science 278, 860-866.
  12. Johnson, L.N. et al. (1996) Cell 85, 149-158.
  13. Delhase, M. et al. (1999) Science 284, 309-313.

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