Product Uses Include
- RhoA biochemistry
- RhoA GTPase assays
- RhoA nucleotide exchange assays
- RhoA binding studies
Material
The human RhoA 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 52 kDa, consisting of the 24 kDa RhoA protein plus a 28 kDa GST tag in the amino-terminus.
Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. GST-RhoA samples are >90% pure.
Biological Activity
The biological activity of RhoA proteins was monitored by measuring the ability of the protein to hydrolyze GTP in the presence and absence of a GTPase-activating protein (RhoGAP) using Cat. # BK055. In the absence of RhoGAP (Cat. # GAS01), GST-RhoA hydrolyzed 0-10% of bound GTP in 5 min at 23ºC, while in the presence of RhoGAP (1:1 RhoGAP : GST-RhoA) 75-80% of bound GTP was hydrolyzed under identical conditions.
