- Other Names
- GTPase Assay, GTPase colorimetric assay
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- Product Citations
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The GTPase colorimetric assay kit employs a 96-well plate format with all of the reagents necessary for measuring GTPase activity. The kit also contains PhosChrome™ (a superior malachite green reagent) which has special additives to prevent backgrounds arising out of non-enzymatic GTP hydrolysis.Product Details
- Kit Contents
PhosChrome™ - 1 bottle, 25mL
Accelerator - 1 vial, 0.5mL
Stabilizer - 1 bottle, 10mL
0.1M MgCl2 - 2 vials, 1.5mL
0.5M Tris pH 7.5 - 1 bottle, 10mL
0.1mM Pi Standard - 1 bottle, 10mL
GTP - 10 vials, lyophilized
96-well plates - 5 plates
Enzyme activity assay
- Recommended Usage
See manual for instructions.
- Application Notes
- High throughput colorimetric enzymatic assay
- Simple to use, scalable, and non-radioactive
- Compatible with any assay buffer
- Stable reagent formulation that ensures long shelf life
- Suppresses non enzymatic backgrounds with acid-labile substrates
- No precipitation, results can be measured over several hours
- Wide linear range, no inhibition of color development by high concentrations of protein
How it works:
Assays are based on the formation of green colored complexes between an inorganic phosphate and PhosChrome™ (an orange colored superior malachite green reagent) under acidic conditions.
(PubMed link indicates BioLegend citation)
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- Biology Area
- Cell Biology, Cell Proliferation and Viability
- Gene ID
- I have 5% DMSO in my assay. Can I use PhosChrome™?
Yes, the reagent is designed for drug screening work and other situations that require DMSO.
- I have phosphate in my enzyme. What can I do?
You can dialyze or desalt the enzyme into a phosphate-free buffer.
- Why do I get a high background when my enzyme definitely has no free Pi?
This is almost certainly caused by inadequate mixing of the Stabilizer. This results in a high background signal because of non-enzymatic decay of ATP substrate. The Stabilizer is added in a relatively small volume (20μL), and the operation of pipetting up and down with a pipette set to 20μL volume may not result in sufficient mixing when the total volume is 270μL. Try pipetting up and down while stirring at the same time. Alternatively, add the Stabilizer with one pipette set at 20μL volume and mix using a larger pipette set to ~150μL volume. This ensures thorough mixing of the Stabilizer solution with minimal effort.