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 5 mg/ml, the protein is in the following buffer: 50 mM Tris pH 7.5, 0.5 mM MgCl2, 50 mM NaCl, 0.5% sucrose, and 0.1% dextran. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent, Cat. # ADV02.
The recombinant protein is 22 kDa, consisting of the Rac1 protein plus a histidine 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. His-Rac1 samples are >90% pure.
Figure 1: His-Rac1 protein purity determination. A 10 µg sample of RC01 (His-Rac1 molecular weight approx. 22 kDa) was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.
The biological activity of RC01 is determined by its ability to exchange nucleotide. This is tested by a pulldown assay using GST-tagged PAK-PBD beads (Cat. # PAK02) and GTPγS (Cat. # BS01) or GDP loaded His-Rac1. The PAK protein is an effector of Rac1, and will specifically bind to the GTP bound form Rac1. Using this assay, the amount of biologically active GTP-bound Rac1 is determined. Stringent quality control ensures that >80% of the His-Rac1 protein produced is capable of binding GTP.
For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at email@example.com
Kulkarni, S., Goll, D. E. and Fox, J. E. (2002). Calpain cleaves RhoA generating a dominant-negative form that inhibits integrin-induced actin filament assembly and cell spreading. J. Biol. Chem. 277, 24435-24441.