Format: 96-well plate

Form: 96 Tests

Detection Method: Colorimetric

Sensitivity: 0.05 mUnits

Assay Range: 6-100 mU/well

Unit Definition: One unit is defined to the amount of enzyme required to transfer 1 nmol phosphate per min to substrate at 30° C.

Assay Time: 3 h

Sample Type: Cell lysates, tissue extracts, purified enzyme

Kit Contents: Streptavidin-Coated 96-Well Plate, Akt Substrate, Biotinylated, Phosphoserine Detection Antibody, Antibody Diluent, Kinase Assay Buffer, 5X ATP/MgCl2 Mix, 100X Inhibitor, Kinase Stop Solution, ELISA 20X Plate Wash Concentrate, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol. Sold under license of U.S. patent 6,441,140. Manufactured by Cell Signaling.

Comments: The K-LISA™ Akt Activity Kit is a rapid, sensitive, ELISA-based activity assay suitable for measuring the kinase activity of purified or partially purified Akt preparations, in vitro Akt inhibitor screening, and for assessing the regulation of Akt in cell signaling. The assay utilizes a biotinylated peptide substrate (GRPRTSSFAEG) that is phosphorylated on the second serine by Akt1, Akt2, Akt3, SGK (Serum Glucocorticoid Kinase), and MSK1. Biotinylated Akt substrate and sample containing Akt are incubated in the presence of ATP in wells of a streptavidin-coated 96-well plate, which allows for phosphorylation and substrate capture in a single step.

Ref.: Liau, S.S., et al. 2006. J. Gastrointest. Surg. 10, 1254. Hill, M.M. et al. 2002. Pharmacol. Ther. 93, 243. Shin, I. et al. 2002. Nat. Med. 8, 1136. Zheng, et al. 2002. J. Biol. Chem. 42, 39379. Brazil, D.P. et al. 2001. Trends Biochem. Sci. 26, 657. Medema, R.H., et al. 2000. Nature 404, 782. Datta, S.R., et al. 1999. Genes Dev. 13, 2905. Franke, F.T., et al. 1997. Cell 88, 435. Alessi, D.R., et al. 1996. EMBO J. 15, 6541.

*Sold under license of U.S. patent 6,441,140 Manufactured by Cell Signaling