IKK beta (IkappaB Kinase beta, IKKb, IKK2)

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). IKK1/a and IKK2/b are phosphorylated by NF-kB-inducing kinase (NIK) (6) and MAP kinase kinase kinase-1 (MEKK1) (7), respectively. Recent studies have shown that IKK2/b, not IKK1/a, is the target proinflammatory stimuli (9). Targeted disruption of IKK2/b gene in mice also results in extensive liver damage from apotopsis and death as embryo (9). Immunoprecipitation: 2ug/ 1x10(6) cells. 2 ug was used to immunoprecipitate IKKb from 1x10(6) cells and the immuno-precipitated protein was detected by western blotting. IKKb was detected as a protein of ~87 kD. \n\nApplications: \nWestern blotting. Whole cell lysate from Daudi, Jurkat, HeLa, or NIH 3T3 cells can be used as positive control and an approximately 87kD band should be detected. It will cross-react with mouse IKKb. Immunoprecipitation, 2 ug of IMG-159 was used to immunoprecipitate IKKb from 1x10(6) cells and the immuno-precipitated protein was detected by western blotting using IMG-129. IKKb was detected as a protein of ~87 kD. \n\nRecommended Dilution: \nWestern blot: 1-2ug/ml\nImmunoprecipitation: 2ug/ 1x10(6) cells\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze 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. \n

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

I3000-24B

Size

100ug

Applications

IP, WB

Hosts

Mouse

Reactivities

Mouse

Form

Supplied as a liquid in PBS, 0.02% sodium azide.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

1. Verma, I.M., Stevenson, J.K., Schwarz, EM., Van Antwerp, D. & Miyamoto, S. Genes. Dev. 9: 2723-2735 (1995). \n2. Verma, I. And Stevenson, J.K. Proc. Nat. Acad. Sci. USA 94: 11758 (1997). \n3. Didonato, J.A., Hayakawa, M., Rothwarf, D.M., Zandi, E., and Karin, M. Nature 388: 548- (1997). \n4. Regnier, C.H., Ho, Y.S., Gao, X., Goeddel, D.V., Cao, Z., and Rothe, M. Cell 90, 373 (1997). \n5. Mercurio, F., Zhu, H., Murray, B.W., Shevchenko, A., Bennett, B.L., Li, J., Young, D.B., Barbosa, M., Mann, M., Manning, A., Rao, A. Science 278: 860 (1997). \n6. Malinin, N.L., Boldin, M.P., Kovalenko, A.V., and Wallach, D. Nature 385: 540-544 (1997). \n7. Lee, F.S., Hagler, J., Chen, J., and Maniatis, T. Cell 88: 213-222 (1997). \n8. Delhase, M., Hayakawa, M., Chen, Y., and Karin, M. Science 284: 309-313 (1999). \n9. Li. Q., Van Antwerp, D., Mercurio, F., Lee, K-F., and Verma, I.M. Science 284: 321-325 (1999).

Additional Info

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). IKK1/a and IKK2/b are phosphorylated by NF-kB-inducing kinase (NIK) (6) and MAP kinase kinase kinase-1 (MEKK1) (7), respectively. Recent studies have shown that IKK2/b, not IKK1/a, is the target proinflammatory stimuli (9). Targeted disruption of IKK2/b gene in mice also results in extensive liver damage from apotopsis and death as embryo (9). Immunoprecipitation: 2ug/ 1x10(6) cells. 2 ug was used to immunoprecipitate IKKb from 1x10(6) cells and the immuno-precipitated protein was detected by western blotting. IKKb was detected as a protein of ~87 kD. \n\nApplications: \nWestern blotting. Whole cell lysate from Daudi, Jurkat, HeLa, or NIH 3T3 cells can be used as positive control and an approximately 87kD band should be detected. It will cross-react with mouse IKKb. Immunoprecipitation, 2 ug of IMG-159 was used to immunoprecipitate IKKb from 1x10(6) cells and the immuno-precipitated protein was detected by western blotting using IMG-129. IKKb was detected as a protein of ~87 kD. \n\nRecommended Dilution: \nWestern blot: 1-2ug/ml\nImmunoprecipitation: 2ug/ 1x10(6) cells\n\nStorage and Stability:\nIt is supplied as Protein G purified IgG fraction. Store at 4oC, stable for 6 months. For long-term storage, store at -20oC. \n