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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|Teckchandani, Anjali M. et al.||c-Cbl regulates migration of v-Abl-transformed NIH 3T3 fibroblasts via Rac1||Experimental cell research||2005||ISSN 0014--4827|
|Patil, Suresh B. et al.||Phosphorylated HSP27 essential for acetylcholine-induced association of RhoA with PKCalpha||American journal of physiology. Gastrointestinal and liver physiology||2004||ISSN 0193--1857|
|Yamashita, Toshihide et al.||The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI||Nature neuroscience||2003||ISSN 1097--6256|
|Lesser, C. F. et al.||Expression of microbial virulence proteins in Saccharomyces cerevisiae models mammalian infection||The EMBO Journal||2001||ISSN 1460--2075|
|Slater, S. J. et al.||Interaction of Protein Kinase C Isozymes with Rho GTPases†||Biochemistry||2001||ISSN 0006-2960|
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