• Study in vitro regulation of RhoA cell signaling mechanisms
• Substrate for identification of RhoA binding proteins

RhoA is a small GTPase that regulates actin cytoskeleton dynamics, primarily promoting the formation of stress fibers and focal adhesions to support cell contractility and adhesion. Through downstream effectors such as ROCK and mDia, RhoA controls actin filament bundling, myosin II activation, and the stabilization of actin structures critical for cell shape and migration.

The constitutively active form of human RhoA protein is produced in a bacterial expression system. The protein has a glutamine to leucine substitution at amino acid 63, creating a constitutively active mutant protein that will not hydrolyze GTP.

• Protein tag: N-terminal His tag for purification
• Molecular weight: approximate mwt 22-25 kDa
• Formulation: supplied as a lyophilized powder
• Composition: when resuspended to 1 mg/ml, the buffer composition is: 10 mM Tris pH 7.5, 10 mM NaCl, 0.1 mM MgCl2, 1.0% sucrose, and 0.2% dextran

Protein purity is assessed by scanning densitometry of Coomassie Blue-stained protein on a 4-20% polyacrylamide gel. Purity was determined to be ≥90% pure.

The biological activity of R6301 is demonstrated by a pull-down assay using GST-tagged Rhotekin RBD beads Cat.# RT02. The Rho Binding Domain (RBD) of the Rho effector protein Rhotekin has a high affinity for GTP-bound Rho. Using this assay, the amount of biologically active GTP-bound RhoA or constitutively active mutant RhoA can be determined. Stringent quality control ensures that >80% of the RhoA protein is pulled down in this assay.