Product Uses Include
- Rap1b biochemistry
- Rap1b GTPase assays
- Rap1b nucleotide exchange assays
- Rap1b binding studies
Material
The human Rap1b 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 approximately 26 kDa, consisting of the Rap1b protein plus a linker and a 6X His tag in the amino-terminus.
Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Rap1b samples are >90% pure.
Biological Activity
The biological activity of Rap1b is determined from its ability to exchange GDP for GTP. EDTA is used to sequester magensium ions from the Rap1b protein, thereby stimulating nucleotide exchange activity. The RhoGEF exchange assay Biochem Kit™ (Cat. # BK100) is used to monitor the exchange
