ADP-Glo Kinase Assay + ACK Kinase Enzyme System
Recombinant human ACK (110-476) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ACK is a tyrosine kinase that binds CDC42Hs in its GTP-bound form and inhibits both the intrinsic and GTPase-activating protein (GAP)-stimulated GTPase activity of CDC42Hs (1). Overexpression of ACK in cancer cell lines of epithelial origin increases cellular motility and invasiveness. In a mouse model, ACK overexpression enhances the ability of a human breast cancer cell line to metastasize to the lung and increased mortality (2). Ligand stimulation of alpha-3 beta-1 integrin leads to activation of ACK which then enhances p130CAS phosphorylation and activation of RAC. 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: ACK, 10ug (Human, recombinant; amino acids 110-476). MW: ~66kDa. Substrate: Poly (4:1 Glu, Tyr) Peptide. Other: Reaction Buffer, DTT, MnCl2. ACK NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/10188/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.