C3 transferase protein

Question 1: What is the difference between the Rho inhibitors CT03 and CT04?

Answer 1: The only difference between these C3 Transferase proteins (Cat. #CT03 and CT04) is that CT04 is covalently linked to a proprietary cell penetrating moiety via a disulfide bond. In this way, CT04 is a much better reagent to use to inhibit Rho activity in living cells.

Question 2: How can I assess whether Rho activity is changing following CT03 treatment?

Answer 2: There are multiple ways to measure changes in Rho activity. If CT03 has been delivered to cells via micro-injection or pinocytic uptake, we recommend examining Rho-mediated stress fiber formation with fluorescently-labeled phalloidin (Cat. #PHDG1, PHDH1, PHDN1, PHDR1). These Acti-stain phalloidins label F-actin stress fibers. Activation of Rho can be directly quantified with one of our activation assays, either the traditional pull-down (Cat. #BK036) or the RhoA G-LISA activation assay (Cat. #BK124). You can also measure CT03&rsquo, s ability to ADP ribosylate native Rho in human platelet extracts in vitro (please see the CT03 datasheet for more details). Briefly, a standard biological assay for monitoring the ADP ribosylation of Rho consists of an in vivo ribosylation reaction followed by non-denaturing gel electophoresis and Western blot analysis using Cytoskeleton&rsquo, s anti-Rho monoclonal antibody (Cat. #ARH03). Stringent quality control ensures that >, 80% of native Rho protein is ADP ribosylated by the recombinant C3 transferase.