IKKe/IKK-i, TBK1

NF-kB (nuclear factor kB) is sequestered in the cytoplasm by IkB family of inhibitory proteins that mask the nuclear localization signal of NF-kB thereby preventing translocation of NF-kB to the nucleus (1). External stimuli such as tumor necrosis factor or other cytokines results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes. Synthesis of IkBa is autoregulated (2). IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g (3-5). Using a subtractive hybridization technique, a novel kinase, IKKi/IKKe has been isolated (6). LPS increases IKKi mRNA level in mouse macrophage cell lines. This protein has significant sequence homology with kinase domains of IKK1/a and IKK-2/b. Overexpression of wild type IKKi in cells phosphorylates Ser32 and Ser36 of IkBa. \n\nApplications: \nSuitable for use in Western Blot. Other applications not tested. \n\nRecommended Dilution: \nWestern blot: 1-3ug/ml Detectis human IKKe in Jurkat, 293 or mouse NIH 3T3 whole cell lysate using 15ug ugs of lysate. Optimal dilutions to be determined by the researcher.\n\nRecommended Positive Control: \n293, Jurkat, or NIH 3T3.\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

I3000-61

Size

100ug

Applications

WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat

Form

Supplied as a liquid in PBS, pH 7.2, 0.05% sodium azide.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG2a,k

References

1. Verma, I.M.,,et al.. Genes. Dev. 9: 2723-2735 (1995). 2.

Additional Info

Recognizes human IKKi/IKKe/TBK1 at ~80kD. Species Cross-reactivity: mouse and rat. Sequence homology: kinase domains of IKK1/a, IKK-2/b and overexpression of wild type IKKi in cells phosphorylates Ser32 and Ser36 of IkBa.