RhoA G-LISA Activation Assay (colorimetric)

On Arrival: 4°C

The kit contains sufficient reagents to perform 96 RhoA activation assays. Since the Rho-GTP affinity wells are supplied as strips and the strips can be broken into smaller pieces, each kit can be used for anywhere from one to multiple assays. The following components are included in the kit:
Rho-GTP affinity wells (12 strips of 8 wells)
Lysis buffer
Binding buffer
Antigen presenting buffer
Wash buffer
Antibody dilution buffer
Anti-RhoA antibody
HRP-labeled secondary antibody
Positive control RhoA protein
Protease inhibitor cocktail (Cat. #PIC02)
Absorbance detection reagents
Precision Red Advanced protein assay reagent (Cat. # ADV02)
Manual with detailed protocols and extensive troubleshooting guide

Rho signaling pathway studies
Rho activation assays with primary cells
Studies of Rho activators and inactivators
Rho activation assays with limited material
High throughput screens for Rho activation

Serum starved Swiss 3T3 cells were stimulated with the Rho activating compound calpeptin and RhoA activation was measured with the G-LISA method (Figures 1 and 2)

Figure 2. Rho activity measured in Swiss 3T3 cells treated with the Cell Permeable Rho Inhibitor (CT04) using the RhoA G-LISA Activation Assay (Cat.# BK124). Serum starved Swiss 3T3 fibroblasts were untreated (no CT04) or treated with 0.20, 0.50 and 2.0 ug/ml of CT04 for 4h in serum free medium at 37C, then activated with 100ug/ml calpeptin for 10min. Cells were then lysed and RhoA activity was measured by the RhoA G-LISA Activation Assay (Cat.# BK124). Note: At 2.0 ug/ml CT04 for 4h results in almost complete (90%) inhibition of RhoA activity.