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.