K-LISA™ Chk1/Chk2 Activity Kit

Format: 96-well plate

Form: 96 Tests

Detection Method: Colorimetric

Sensitivity: 25 pg (0.018 mU) purified Chk1

Assay Range: 25-725 µU (Chk1) and 260 µU - 26 mU (Chk2)

Assay Time: 3 h

Sample Type: Cell lysates, tissue extracts, purified enzymes

Kit Contents: Biotinylated Chk1 Substrate, Phosphoserine Detector Antibody, Streptavidin Coated 96-Well Plate Strips, HRP-Conjugated Secondary Antibody, Serine/Threonine Kinase Assay Buffer, ATP/MgCl2, Chk1 Inhibitor, Wash Buffer, TMB Substrate, Stop Solution, Plate Sealers, and a user protocol. Purified Chk1 is available separately (Cat. No. 220479).

Comments: The K-LISA™ Checkpoint Activity Kit is a rapid, sensitive, ELISA-based activity assay suitable for measuring the kinase activity of purified or partially purified Chk1 and Chk2 preparations, in vitro Chk1 and Chk2 inhibitor screening, and for assessing the regulation of Chk1 and Chk2 in cell signaling. The assay utilizes a biotinylated peptide substrate (KKKVSRSGLYRSPSMPENLNRPR) that is phosphorylated on the third serine by Chk1 and Chk2. Biotinylated Chk substrate and sample containing Chk1 or Chk2 are incubated in the presence of ATP in wells of the streptavidin-coated 96-well plate, which allows for phosphorylation and substrate capture in a single step.

Ref.: Bartek, J., and Lukas, J. 2003. Cancer Cell 3, 421. O'Neill T., et al. 2002. J Biol. Chem. 277, 16102. Jackson, J.R., et al. 2000. Cancer Res. 60, 566. Liu, Q., et al. 2000. Genes Dev. 14, 1448.