Product Uses Include：

Precipitation of active Rho from cell lysates

Inhibition of RhoA/B/C activity in vitro

Blocking of RhoA/B/C function in vivo by microinjection

Material：
       The Rho binding domain (RBD) of the human Rhotekin protein has been expressed as a GST-fusion protein in E. coli. This protein binds binds specifically to GTP-bound, and not GDP-bound, RhoA, RhoB and RhoC proteins. The domain can therefore be used to specifically precipitate active, GTP-bound Rho proteins as well as to specifically block the activity of RhoA/B/C in vitro and in vivo.

       The GST-Rhotekin-RBD contains residues 7-89 of Rhotekin. This region includes the sequences required for the high affinity interaction with GTP-Rho.

The protein is supplied as 500 μg of lyophilized product and the protein is also available in a bead bound format (Cat. # RT02). This product is used in our RhoA pulldown activation assay Biochem Kit? (Cat. # BK036)

Purity：
       Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% SDS polyacrylamide gel. GST-Rhotekin-RBD protein is >90% pure (see Figure 1).

       Figure 1:  GST-Rhotekin-RBD protein purity determination. A 10 μg sample of RT01 was separated by electrophoresis in a 12% SDS-PAGE system and stained with Coomassie Blue.

Biological Activity：
       The Rhotekin-RBD protein specifically recognizes and binds to the active, GTP-bound, form of RhoA, RhoB or RhoC protein. It has a much lower affinity for the inactive, GDP-bound, form of RhoA/B/C. Biological activity of Rhotekin-RBD protein is therefore determined by its selectivity for GTP-RhoA protein.