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 | -20°C |  | Dry ice |  | Multiple Toxicity Values, refer to MSDS | | Note: Store and Ship conditions may differ. | | See Key |
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| | K-LISA™ Aurora Kinase Activity Kit |
|  | Cat. No. CBA051 |
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| Format: 96-well plate | | Form: 96 Tests | | Detection Method: Colorimetric | | Specificity: Aurora A, Aurora B | | Sensitivity: 16 ng for Aurora A; 11 ng for Aurora B | | Assay Range: 20-1000 ng | | Sample Type: Purified enzyme or enzyme immunoprecipitated from cell lysates | | Kit Contents: Streptavidin-Coated 96-Well Plate, Aurora Substrate, Biotinylated, Anti-Phospho Histone H3a 5X Kinase Assay Buffer, 20 mM ATP, 100X Inhibitor, Kinase Stop Solution, ELISA 20X Plate Wash Concentrate, Antibody Diluent, HRP-Antibody conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol | | Comments: An ELISA-based activity assay for measuring the kinase activity of purified or partially purified Aurora A and Aurora B, in vitro Aurora A and Aurora B inhibitor screening, and for assessing the regulation of Aurora A and Aurora B in cell signaling. | | Ref.: Meraldi, P., et al. 2004. Curr. Opin. Genet. Dev. 14, 29. Ewart-Toland, A., 2003.Nat. Genet. 34, 403. Kufer, T.A., et al. 2002. J. Cell Biol. 158, 617. Hsu, J.Y., et al. 2000. Cell 102, 279. |
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Activity of Aurora A Immunoprecipitated From HeLa Cell Lysate.. HeLa cells were treated with 50 ng/ml nocodazole (Cat. No. 487928) for 16 h and cell lysate was prepared using 0.5 ml PhosphoSafe™ Extraction Buffer (Cat. No. 71296) for each 10 cm cell culture dish, according to the recommended protocol. Cell lysates were either used immediately or stored at -70°C until use. Equal amounts of total protein (500 µg) were used to immunoprecipitate Aurora A using Anti-Aurora A, Rabbit pAb (Cat. No. PC742). Activity was determined using protocol B as outlined in the Detailed Protocol section above.
Activity of Purified Aurora A and Aurora B. The activity of Aurora A, Human, Recombinant, His•Tag® (Cat. No. 481413) (1.66-1660 ng) and Aurora B (0.5-500 ng) was determined using protocol A as outlined in the Detailed Protocol above. Assay range: 20-1000 ng.
Staurosporine Inhibition of Aurora A Activity.. The activity of Aurora A, Human, Recombinant, His•Tag® (Cat. No. 481413) (200 ng) was determined as in Figure 1 in the presence of increasing Staurosporine concentrations.
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