ADP-Glo Kinase Assay + PAK1/CDC42 Kinase Enzyme System
Recombinant full-length human PAK1 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag and combined with CDC42 at a ratio of 1:4 (PAK1:CDC42) in vitro. PAK1 is a member of the p21-activated kinases (PAKs), which have been implicated in the regulation of cell morphology, motility and transformation. These serine/threonine kinases are activated by and are effectors of small GTPases, CDC42 and RAC. PAK1 is one of the Group I PAKs, which also include PAK2 and PAK3 (1). PAK1 is a key regulator of the actin cytoskeleton, adhesion and cell motility. Inactive dimeric PAK1 is mainly cytosolic and interaction with the activators Cdc42-GTP and Rac1-GTP stimulates the kinase at the sites of cellular protrusions forming adhesions to the extracellular matrix (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: PAK1/CDC42, 10ug (Human, recombinant full-length). MW: ~96kDa (PAK1). Substrate: PAKtide (RRRLSFAEPG). Other: Reaction Buffer, DTT, GTP Solution. PAK1/CDC42 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/5058/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.