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Encompassing Amino Acids: 227-524
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). PAK2 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 20, 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: PAK proteins is a family of serine/threonine kinases that serve as targets for the small GTP binding proteins, CDC42 and RAC1, and have been implicated in a wide range of biological activities. The p21 activated kinases (PAK) are critical effectors that link Rho GTPases to cytoskeleton reorganization and nuclear signaling. PAK2 is activated by proteolytic cleavage during caspase-mediated apoptosis, and may play a role in regulating the apoptotic events in the dying cell. The CDC42 and RAC1 induce autophosphorylation of PAK2, which stimulates sustained phosphorylation of other substrates.
Description: Human PAK2 (GenBank Accession No. NM_002577), a.a. 227-524, MW=33.5 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_002577
Purity: ≥95%
Storage Stability: At least 6 months at -80°C.
Tags: none
Uniprot: Q13177
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Coniglio,S.J., et al., Mol. Cell. Biol. 28 (12), 4162-4172 (2008).
2. Koh,W., et al., J. Cell. Sci. 121 (PT 7), 989- 1001 (2008).
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