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Question 1: Can the Rho inhibitor CT04 be used with cells growing in culture?
Answer 1: Yes, Cytoskeleton modified the exoenzyme C3 Transferase from Clostridium botulinum to be cell permeable (Cat. #CT04) for the specific purpose of inhibiting Rho in living cells in as little as 2 hours.
The exoenzyme C3 Transferase from Clostridium botulinum is commonly used to selectively inactivate the GTPases RhoA, RhoB, and RhoC, both in vivo and in vitro. C3 Transferase inhibits Rho proteins by ADP-ribosylation on asparagine 41 in the effector binding domain of the GTPase. A major limitation of the standard C3 Transferase for use in in vivo applications is that this protein is not cell permeable.
Cytoskeleton&rsquo, s cell permeable Rho inhibitor consists of highly purified C3 Transferase covalently linked to a proprietary cell penetrating moiety via a disulfide bond. The cell penetrating moiety allows rapid and efficient transport through the plasma membrane. Once in the cytosol, the cell penetrating moiety is released, thereby allowing C3 Transferase to freely diffuse intracellularly and inactive RhoA, RhoB, and RhoC, but not related GTPases such as Cdc42 or Rac1.
Question 2: How can I assess whether Rho activity is changing in my cells following CT04 treatment?
Answer 2: There are multiple ways to measure changes in Rho activity. To visualize a change in Rho activity, 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).
PurityThe Exoenzyme C3 Transferase from Clostridium botulinum has been produced in a bacterial expression system. The recombinant protein contains six histidine residues at its amino terminus (His tag), has a molecular weight of approximately 24 kDa, and is greater than 90% pure (see Cat. # CT03 for gel purity). To make the purified C3 Transferase protein cell permeable, a proprietary cell penetrating moiety has been linked via a reversible disulfide bond. Cell Permeable Rho Inhibitor is supplied as a white lyophilized powder.
Hinweis: Die dargestellten Informationen und Dokumente (Bedienungsanleitung, Produktdatenblatt, Sicherheitsdatenblatt und Analysezertifikat) entsprechen unserem letzten Update und sollten lediglich der Orientierung dienen. Wir übernehmen keine Garantie für die Aktualität. Für spezifische Anforderungen bitten wir Sie, uns eine Anfrage zu stellen.
Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.
CT04-C.pdf
CT04-C-datasheet.pdf
CT04-C-safety-datasheet.pdf
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