ADP-Glo Kinase Assay + CHK2 Kinase Enzyme System
Recombinant full-length human CHK2 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. CHK2 is rapidly phosphorylated and activated in response to replication blocks and DNA damage; the response to DNA damage occurs in an ataxia telangiectasia mutated (ATM)-dependent manner (1). Expression of wild-type CHK2 leads to increased p53 stabilization after DNA damage, whereas expression of a dominant-negative CHK2 mutant abrogated both phosphorylation of p53 on Ser-20 and p53 stabilization (2). ADP-Glo Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP. Kinase Enzyme System contains: Kinase: CHK2, 10ug (Human, recombinant full-length). MW: ~88kDa. Substrate: Chktide (KKKVSRSGLYRSPSMPENLNRPR); derived from human CDC25C protein isoform A (amino acids 205-225). Other: Reaction Buffer, DTT. CHK2 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/11200/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.