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, 10μg (Human, recombinant full-length). MW: ~96kDa (PAK1). Substrate: PAKtide (RRRLSFAEPG). Other: Reaction Buffer, DTT, GTP Solution.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Jaffer, Z.M. et al. (2002) p21-activated kinases: three more join the Pak. Int J Biochem Cell Biol. 34, 713–7.
2.Parrini, M.C. et al. (2005) Spatiotemporal regulation of the Pak1 kinase. Biochem Soc Trans. 33, 646–8.
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