Product Uses Include：

Precipitation of active Rac and Cdc42 from cell lysates

Inhibition of Rac and Cdc42 activity in vitro

Blocking of Rac and Cdc42 function in vivo by microinjection

Material：
       The Rac/Cdc42 (p21) binding domain (PBD) of the human p21 activated kinase 1 protein (PAK) protein has been expressed as a GST-fusion protein in E. coli. This protein binds binds specifically to GTP-bound, and not GDP-bound, Rac and Cdc42 proteins. The domain can therefore be used to specifically precipitate active, GTP-bound Rac and Cdc42 as well as to specifically block the activity of Rac and Cdc42 in vitro and in vivo.

       The GST-PAK-PBD contains residues 67-150 of PAK. This region includes the highly conserved CRIB motif plus sequences required for the high affinity interaction with GTP-Rac and GTP-Cdc42.

       The protein is supplied as 250 µg of lyophilized product and the protein is also available in a bead bound format (Cat. # PAK02). This product is also used in our Cdc42 and Rac activation assay Biochem Kits™ (Cat. # BK034 and BK035, repectively)

Purity：
       Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% SDS polyacrylamide gel. GST-PAK-PBD protein is >80% pure (see Figure 1).

        Figure 1: GST-PAK-PBD protein purity determination. A 10 µg sample of PAK01 was separated by electrophoresis in a 12% SDS-PAGE system and stained with Coomassie Blue.

Biological Activity：
       The PAK-PBD protein specifically recognizes and binds to the active, GTP-bound, form of Rac and Cdc42. Biological activity of PAK-PBD protein is therefore tested by its selectivity for GTPγS loaded Rac and Cdc42 proteins