DESCRIPTION
Para-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.
phosphatase
p-Nitrophenyl phosphate ------> p-nitrophenol
+ phosphate
This homogeneous "mix-and-measure" assay involves simply adding a single
reagent to the phosphatase and measuring the product formation using any
absorbance reader. The assay can be conveniently performed in cuvettes,
tubes or multi-well plates at either room temperature or 37°C. In addition,
the reagents are compatible with ELISA assays in which alkaline phosphatase
conjugated secondary antibody is used. This kit is well adapted for a number
of applications. For example, it can be utilized for direct characterization
of enzyme activity and for assay condition optimization. It can also be
applied for research diagnosis of diseases that are associated with
increased levels of alkaline phosphatase. Typical diseases include liver
disease, bone disease, Hodgkin's disease, congestive heart failure,
Fanconi's syndrome, hyperparathyroidism, intestinal disease and abdominal
bacterial infections. Moreover, it can be used to characterize phosphatase
inhibitors through high-throughput screening.
The kit reagents have been optimized for long shelf-life and maximum
reproducibility. The reagents are compatible with all liquid handling
systems and bulk reagents are available for high-throughput screening of
phosphatase inhibitors.
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