Cell Signaling Technology

Product Pathways - NF-kB Signaling

Phospho-IKKγ (Ser376) Antibody #2689

Applications Reactivity Sensitivity MW (kDa) Source
W H Endogenous 50 Rabbit

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

Protocols

Specificity / Sensitivity

Phospho-IKKγ (Ser376) Antibody detects endogenous levels of IKKγ protein only when phosphorylated at Ser376.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser376 of human IKKγ protein. Antibodies are purified by protein A and peptide affinity chromatography.

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 (upper) or IKKγ Antibody #2685 (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 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 mouse, mutation of the orthologous residue (Ser369) to alanine leads to enhanced IKKγ-mediated stimulation of IKKβ kinase activity (15).

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  14. Carter, R. S. et al. (2003) J. Biol. Chem. 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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