Product Uses Include
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.
For other forms of RhoA as well as many other purified small G-proteins, see our main small G-protein product page.
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. GST-RhoA samples are >90% pure.
Figure 1: GST-RhoA protein purity determination. A 20 µg sample of RHG01 (GST-RhoA molecular weight approx. 52 kDa) was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.
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.
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