Cell Signaling Technology

Product Pathways - NF-kappaB Signaling

IKKγ (DA10-12) Mouse mAb #2695

Applications Reactivity Sensitivity MW (kDa) Source Isotype
W H M R Endogenous 50 Mouse IgG1

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

Specificity / Sensitivity

IKKgamma (DA10-12) Mouse mAb detects endogenous levels of total IKKgamma protein.

Source / Purification

Monoclonal antibody is produced by immunizing mice with full length human GST-IKKgamma protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, NIH/3T3 and C6 cells using IKKgamma (DA10-12) Mouse mAb (upper) or IKKgamma Antibody #2685 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with TNF-alpha (20 ng/ml) and calyculin A #9902 (50 nM), using Phospho-IKKgamma (Ser376) Antibody #2689 (top) or IKKgamma (DA10-12) Mouse mAb (bottom).

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).

Activation of the NF-kappaB pathway by the T-cell lymphotrophic virus Tax protein or by TNF-alpha treatment leads to IKKbeta-dependent phosphorylation of human IKKgamma, primarily at Ser376 (14). In mice, mutation of the orthologous residue (Ser369) to alanine leads to enhanced IKKgamma-mediated stimlulation of IKKbeta kinase activity (15).

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  13. Delhase, M. et al. (1999) Science 284, 309-313.
  14. Carter, R. S. et al. (2003) 278, 19642-19648.
  15. Prajapati, S. and Gaynor, R.B. (2002) J. Biol. Chem. 277, 24331-24339.

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