Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - NF-kB Signaling

IKKε (D61F9) XP® Rabbit mAb #3416

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F M R Endogenous 80 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  M=Mouse  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

IKKε (D61F9) XP® Rabbit mAb detects endogenous levels of total IKKε protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxyl terminus of mouse IKKε.

Western Blotting

Western Blotting

Western blot analysis of extracts from Raw 264.7 and KNRK cell lines using IKKε (D61F9) XP® Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells (blue) and Raw 264.7 cells (green) using IKKε (D61F9) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of RAW 264.7 (left) and C2C12 (right) cells using IKKepsilon (D61F9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).


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).

Recently, two homologs of IKKα and IKKβ have been described, called IKKε (also known as IKK-i) and TBK-1 (also known as T2K or NAK), and activation of either of these kinases results in NF-κB activation. IKKε contains the kinase domain in its amino terminus, which shares 30% identity to that of IKKα or IKKβ. IKKε is expressed mainly in immune cells, and may play a special role in the immune response (14-18).

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  14. Shimada, T. et al. (1999) Int Immunol 11, 1357-62.
  15. Peters, R.T. et al. (2000) Mol Cell 5, 513-22.
  16. Tojima, Y. et al. (2000) Nature 404, 778-82.
  17. Bonnard, M. et al. (2000) EMBO J 19, 4976-85.
  18. Peters, R.T. and Maniatis, T. (2001) Biochim Biophys Acta 1471, M57-62.

Application References

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Companion Products

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

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