Cdc42-His Protein: Wild type (CD01-GRP)

Product Uses Include

• Cdc42 biochemistry
• Cdc42 GTPase assays
• Cdc42 nucleotide exchange assays
• Cdc42 binding studies

Material
The human Cdc42 protein has been produced in a bacterial expression system. The protein is supplied as a lyophilized powder. When it is reconstituted in distilled water to 1 mg/ml, the protein is in the following buffer: 2 mM Tris pH 7.6, 0.5 mM MgCl₂, 0.5% sucrose, 0.1% dextran. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent, Cat. # ADV02.

The recombinant protein is 22 kDa, consisting of the 22 kDa Cdc42 protein plus a 6 amino acid histidine tag in the amino-terminus.

Purity

Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. His-Cdc42 samples are >90% pure.

Biological Activity

The biological activity of CD01 is determined by its ability to exchange nucleotide. This is tested by a pulldown assay using GST-tagged PAK-1 PBD beads (Cat. # PAK02) and GTPγS (Cat. # BS01) or GDP loaded His-Cdc42. The PAK (p21 activated kinase) protein is an effector of Cdc42, and will specifically bind to the GTP bound form Cdc42. Using this assay, the amount of biologically active GTP-bound Cdc42 is determined. Stringent quality control ensures that >70% of the Cdc42 protein produced is capable of binding GTP.