Cell Signaling Technology

Product Pathways - NF-kappaB 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.

Specificity / Sensitivity

Phospho-IKKgamma (Ser376) Antibody detects IKKgamma only when phosphorylated at serine 376.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser376 of human IKKgamma. 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-alpha (20 ng/ml) and calyculin A #9902 (50 nM) using Phospho-IKKgamma (Ser376) Antibody (top) or IKKgamma Antibody #2685 (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 mouse, mutation of the orthologous residue (Ser369) to alanine leads to enhanced IKKgamma-mediated stimlulation of IKKbeta kinase activity (15).

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  15. Prajapati, S. and Gaynor, R.B. (2002) J. Biol. Chem. 277, 24331-24339.

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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