Cell Signaling Technology

Product Pathways - NF-kB Signaling

IKKγ (DA10-12) Mouse mAb #2695

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

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

Protocols

Specificity / Sensitivity

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

Source / Purification

Monoclonal antibody is produced by immunizing animals with full length human GST-IKKγ protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with TNF-α (20 ng/ml) and Calyculin A #9902 (50 nM), using Phospho-IKKγ (Ser376) Antibody #2689 (top) or IKKγ (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 and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (Ser176 and Ser180 in IKKα), which causes conformational changes, resulting in kinase activation (10-13).

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

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  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.
  14. Carter, R. S. et al. (2003) 278, 19642-19648.
  15. Prajapati, S. and Gaynor, R.B. (2002) J. Biol. Chem. 277, 24331-24339.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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