Product Uses Include
Actin binding studies
Actin polymerization studies
Material
Profilin protein has been expressed in E.coli via chromatography without a tag. Thus the protein does not contian a 6xHis or other tag. The protein is supplied as a lyophilized powder. When resuspended in nanopure water, the profilin will be in the following buffer: 10 mM Tris pH 8.0, 1 mM EDTA, 1 mM DTT, 5% (w/v) sucrose and 1% (w/v) dextran.
The lyophilized protein is stable at 4°C desiccated (<10% humidity) for up to 1 year.
Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Purity is >95% (see figure 1).
Figure 1. Profilin purity determination. A 10 μg sample of PR02 (profilin molecular weight approx. 15 kDa) was run on an SDS-PAGE gel and stained with coomassie blue. Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat.# ADV02).
Biological activity
Profilin is tested for biological activity by a binding assay with actin. Profilin is bound to poly-L-proline sepharose beads and incubated with G-actin. The profilin beads bind >90% of the available actin by this procedure.