Introduction

The G-switch™ line of small G-protein tools has been developed with an emphasis on creating highly potent reagents that target endogenous Rho family proteins and pathways. In contrast to methods that rely on over-expression or knockdown of target proteins (e.g., DNA transfection of dominant-negative or constitutively-active Rho mutants, RNAi knockdown), G-switch™ reagents act rapidly on the endogenous target protein (in minutes to hours, depending on the product), thereby optimizing the chance of generating a more physiologically relevant response. The G-switch™ product line includes reagents that directly and indirectly modulate Rho family signal transduction, thereby offering a wide range of mechanistic tools to study these critical cellular functions. See Cytoskeleton's web site for the latest G-switch™ information.

Product Uses Include

  • Rho pathway studies in tissue culture cells
  • Study effects of altered Rho pathway on other connected pathways
  • An active control when testing other substances
ProductDescription

Manual

MSDS

Price & Order 

Rho Activator II

Cell permeable, specific for Rho, recommended for use in Rho pathway studies requiring a direct activator of Rho

pdf CN03

Rho Inhibitor I

Cell permeable C3 transferase, specific for Rho, inhibits cellular Rho within 2-4h, recommended for use in Rho pathway studies requiring a rapid and direct inhibitor of RhopdfpdfCT04

Rho/Rac/Cdc42 Activator I

Cell permeable, specific for Rho family, recommended for use in Rho pathway studies requiring a direct activator of Rho family proteins

pdfpdfCN04

Rho Activator I

Inhibits SHP-2 which indirectly activates Rho, recommended for use in Rho pathway studies requiring an indirect activator of Rho

pdfpdfCN01

Rac/Cdc42 Activator II

Binds EGF receptor and activates several pathways, including indirect activation of Rho family proteins, recommended for use in Rho pathway studies requiring an indirect activator of Rac/Cdc42

pdfpdf CN02
Rho Pathway Inhibitor I
ROCK inhibitor, Y-27632, acts downstream of Rho by inhibiting the Rho effector kinases ROCK I and ROCK II, recommended for Rho pathway studies.pdfpdfCN06
C3 Transferase Protein

Non-cell permeable C3 transferase, this product is taken up by cells through pinocytosis over 12-24h, specific for Rho, recommended for use in Rho pathway studies requiring a direct inhibitor of Rho that acts over a longer time period than the cell permeable version of this enzyme.

pdfpdfCT03

 

Specification of Activators:

Activator

Advantage

Disadvantages

Window of activation
(Time after addition of reagent)

Serum compatible

G-switch™
Rho Activator II
(Cat# CN03)

Type: Direct
Class: Selective
Targets: Rho

 

The most robust activator for Rho available

Sustained activation of Rho GTPases for >2hr

Eliminates Ras, PI3K and PLC concomitant activation

Most suitable activator for studying specific signal transduction pathways downstream of the Rho GTPases

Cannot be used to study GAPs

C3 exoenzyme (Cat# CT04) will not inhibit CN03 activated small G-proteins

2 - 12 h, no signal after 24 h*.

YES

G-switch™
Rho/Rac/Cdc42 Activator I
(Cat# CN04)

Type: Direct,
Class:Non-selective
Targets:Rho,Rac,Cdc42

The most robust activator for Rac and Cdc42 available, second only to CN03 for Rho activation

Sustained activation of Rho, Rac and Cdc42 for >2hr

Eliminates Ras, PI3K and PLC concomitant activation

Excellent activator for studying signal transduction pathways downstream of Rho, Rac and Cdc42

Cannot be used to study GAPs

C3 exoenzyme (Cat# CT04) will not inhibit CN04 activated small G-proteins

Rho; 2 - 12 h, no signal after 24 h*.

Rac; 2 - 24 h, no signal after 48 h.

Cdc42; 2 - 24 h, no signal after 48 h.

YES

Rac/Cdc42 Activator
(Cat# CN02)

Type: Indirect,
Class: Non-selective
Targets:Rho,Rac,Cdc42

Physiological stimulus

Useful in studying pathway between cell surface and small G-proteins

Very transient activation

Ras, PI3K and PLC also activated

1 - 3 min, no signal after 5 min.

YES

Rho Activator I
(Cat# CN01)

Type: Indirect
Class: Selective
Target:Rho

More sustained Rho activation than EGF (Cat# CN02)

Eliminates Ras, PI3K and PLC concomitant activation

Also inhibits calpain

Inhibition of SHP-2 may have other biological consequences

Requires DMSO vehicle

10 - 30 min, no signal after 120 min.

YES

*variable signal after 24h (1)

References

1. Boyer L, Turchi L, Desnues B, Doye A, Ponzio G, Mege JL, Yamashita M, Zhang YE, Bertoglio J, Flatau G, Boquet P, Lemichez E. CNF1-induced ubiquitylation and proteasome destruction of activated RhoA is impaired in Smurf1-/- cells. Mol Biol Cell. 2006 Jun;17(6):2489-97