Product Uses Include
- Study of GEF activity with different GTPases.
- Identification of Dbs binding proteins
- Study of Dbs function in vivo by the introduction of His-Dbs into live cells
- Postive control for other RhoGEF studies.
MaterialThe Guanine nucleotide Exchange Factor domain (GEF domain, also known as the DH/PH tandem domain) of the human RhoGEF Dbs (hDbs) has been produced in a bacterial expression system with a 6xHis tag at its amino terminus. Dbs is an efficient GEF for the RhoA and Cdc42 GTPases and it shows weak GEF activity towards Rac1.
The molecular weight of this His-hDbs DH/PH protein is approximately 40 kDa. The protein is supplied as a lyophilized powder. When reconstituted to 2 mg/ml, the protein is in the following buffer: 20 mM Tris pH 7.5, 0.5 mM MgCl2, 0.5% sucrose and 0.1% dextran. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent (Cat. # ADV02).
PurityPurity is determined by scanning densitometry of proteins run on SDS-PAGE gels. Samples are >80% pure.
Figure 1: His-hDbs DH/PH protein purity determination. A 10 µg sample of GE01 was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.
Biological ActivityThe biological activity of the hDbs DH/PH was tested by measuring the ablility of the protein to catalyze the exchange of GDP for mant-GTP on Cdc42 and RhoA (Cat. # CD01 and RH01, respectively) using the RhoGEF exchange assay kit (Cat. # BK100). See Figure 2.
Figure 2: hDbs DH/PH stimulates nuleotide exchange on Cdc42 as determined by BK100
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 tservice@cytoskeleton.com
H. Jeon et al. (2011). Phospholipase D2 induces stress fiber formation through mediating nucleotide exchange for RhoA. Cellular Signalling. v 23, pp 1320-1326.
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