Application
- For quantitative determination of phosphatase enzyme activity.
Key Features
- High sensitivity and wide linear range. The detection limit is generally 3 ng phosphatase or below.
- Homogeneous and simple procedure. No wash or reagent transfer steps are involved. The assay can be completed within 30 minutes.
- Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments.
Method
- OD405nm
Samples
- Protein phosphatases
Species
- All
Procedure
- 30 min
Size
- 500 tests
Detection Limit
- 3 ng
Shelf Life
- 12 months
More Details
- p-nitrophenyl phosphate (pNPP) is a chromogenic substrate for most phosphatases such as alkaline phosphatases, acid phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The reaction yields para-nitrophenol, which becomes an intense yellow soluble product under alkaline conditions and can be conveniently measured at 405 nm on a spectrophotometer. This homogeneous “mix-and-measure” assay involves simply adding a single reagent to the phosphatase and measuring the product formation on an absorbance reader. The assay can be conveniently performed in cuvettes, tubes or multi-well plates at either room temperature or 37°