This nonradioactive colorimetric assay kit contains all the necessary reagents for measuring enzyme activity (everything included in the PiColorlock™ reagent and more) and is ideal for high throughput drug screening.
The kits contain PiColorlock™, a nonradioactive, superior phosphate detection reagent.
Key features of PiColorLock™ include:
- Colorimetric Assay – Competitor assays are radioactive
- Special additive ensures low backgrounds with acid labile substrates
- Unparalleled stability of phosphate–dye complexes
- Reagent is compatible with almost any assay buffer
- No inhibition of color development by high concentrations of protein
- Stable reagent formulation – Long shelf life.
Colorimetric assays for GTPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions.
Competitor assays are beset with several problems including reagent precipitation. The high stability of PiColorLock™ ensures high stability of the colored dye–phosphate complexes (green color – see below).
Components in the two plate GTP assay kit (five plate kit amounts in brackets)
Store at 4°C:
- 1 x 10ml of PiColorLock™* (1 x 25ml)
- 1 x 0.25ml of Accelerator (1 x 0.5ml)
- 1 x 5ml of Stabiliser (1 x 10ml)
- 1 x 1.5ml of 0.1M MgCl2 (2 x 1.5ml)
- 1 x 5ml of 0.5M Tris pH 7.5 (1 x 10ml)
- 1 x 5ml of 0.1mM Pi standard (1 x 10ml).
*Take care, the solution is very acidic.
Store at -20 or -70oC:
- 4 vials of lyophilized GTP (10 vials)
- Store at room temperature
- 2 x 96 well plates (5 plates).