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Encompassing Amino Acids: 248-545
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Analysis of enzymatic activity was performed according to Zlyte assay protocol (Invitrogen). PAK1 was incubated in a buffer containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 200 µM ATP, 0.01% Brij-35, and 2 µM substrate (SER/THR 14, Invitrogen) at room temperature for 1 hour. Developer solution was added to reaction for 1 hour at RT. Fluorescence was detected using λexc=460±40 nm and λem=528±20 nm
Background: 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 belongs to the Group I PAKs which also includes PAK2 and PAK3. 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.
Description: Human PAK1 (GenBank Accession No. NM_002576.4), a.a. 248-545, MW=33.6 kDa, expressed in E. coli.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8, 150 mM NaCl, 10% glycerol, 5 mM DTT
Genbank: NM_002576.4
Purity: ≥95%
Storage Stability: At least 6 months at -80°C.
Tags: none
Uniprot: Q13153
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Sheehan,K.A., et al., Am. J. Physiol., Cell Physiol. 296 (1), C47-C58 (2009).
2. Reddy,S.D., et al., Cancer Res. 68 (20), 8195- 8200 (2008).