Product Uses Include
The human Rac1 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 50 kDa, consisting of the 22 kDa Rac1 protein plus a 28 kDa GST tag in the amino-terminus.
For other forms of Rac1 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-Rac1 samples are >90% pure.
Figure 1: GST-Rac1 protein purity determination. A 20 µg sample of RCG01 (GST-Rac1 molecular weight approx. 50 kDa) was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.
The biological activity of GST-Rac1 protein 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-Rac1 hydrolyzed 40-50% of bound GTP in 5 min at 23ºC, while in the presence of RhoGAP (1:1 RhoGAP : GST-Rac1) 75-80% of bound GTP was hydrolyzed under identical conditions.
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Grabham, P. W., Reznik, B. and Goldberg, D. J. (2003). Microtubule and Rac 1-dependent F-actin in growth cones. J. Cell Sci. 116, 3739-3748.
Li, X., Bu, X., Lu, B., Avraham, H., Flavell, R. A. and Lim, B. (2002). The hematopoiesis-specific GTP-binding protein RhoH is GTPase deficient and modulates activities of other Rho GTPases by an inhibitory function. Mol. Cell. Biol. 22, 1158-1171.
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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.