• Identify GTPase Activating Proteins (GAPs) targeting RhoA, Cdc42, Rac1, or H-Ras: in vitro assay using purified proteins, not suitable for use with cell lysates
• Biochemical characterization of GAP activity
• Identify GTPase/GAP inhibitors in a high-throughput screen format

Kit contents (20 assays for each GTPase)

• Reaction buffer
• P50 RhoGAP domain protein
• RhoA, His tagged protein
• Rac1, His-tagged protein
• Cdc42, His-tagged protein
• Ras, His-tagged protein
• GTP
• CytoPhos reagent
• 96-well plate

Equipment & materials required

• 96-well plate spectrophotometer, temperature-regulated (650 nm absorbance wavelength)

This assay employs an end-point colorimetric absorbance method to track GTP hydrolysis by small GTPases. GAP proteins enhance GTPase activity, intensifying the final color shift.

Key characteristics

• Detection method: Colorimetric absorbance at 650 nm
• Timeframe: Approximately 2 hours