Cat. # RHG01
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.
GST-RhoA protein is also available in a bead bound format, see Cat. # BX01. For other forms of RhoA as well as many other purified small G-proteins, see our main small G-protein product page.
Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. GST-RhoA samples are >90% pure.
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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. |
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.
Examples of publications where this product was used:
Lesser, C. F. and Miller, S. I. (2001). Expression of microbial virulence proteins in Saccharomyces cerevisiae models mammalian infection. EMBO J. 20, 1840-1849.
Patil, S. B., Pawar, M. D. and Bitar, K. N. (2004). Phosphorylated HSP27 essential for acetylcholine-induced association of RhoA with PKCα. Am. J. Physiol. 286, G635-644.
Slater, S. J., Seiz, J. L., Stagliano, B. A. and Stubbs, C. D. (2001). Interaction of protein kinase C isozymes with Rho GTPases. Biochemistry 40, 4437-4445.
Teckchandani, A. M., Panetti, T. S. and Tsygankov, A. Y. (2005). c-Cbl regulates migration of v-Abl-transformed NIH 3T3 fibroblasts via Rac1. Exp. Cell Res. 307, 247-258.
Yamashita, T. and Tohyama, M. (2003). The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI. Nat. Neurosci. 6, 461-467.
| Product description | Cat. # | Amount | Price & Order |
| Wild type RhoA protein, GST-tagged | RHG01-C | 8 x 25 µg |
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