Protein, alkaline, and acid phosphatases are all measured using the non-specific, non-proteinaceous substrate known as p-Nitrophenyl Phosphate (PNPP). So, it is an ideal reaction for this laboratory assay.
<h3>What is p-nitrophenolphosphate ?</h3>
In ELISA and traditional spectrophotometric experiments, para-Nitrophenylphosphate (pNPP) is a non-proteinaceous chromogenic substrate for alkaline and acid phosphatases.
The hydrolysis of pNPP by phosphatases releases inorganic phosphate and the conjugate base of para-nitrophenol (pNP).
The resultant phenolate has a maximum absorbance at 405 nm and is yellow in colour.
Alkaline phosphatase (AP) and protein tyrosine phosphatase are two phosphatases that can be measured using this characteristic (PTP).
The material needs to be stored in a dark, shielded area because it is light-sensitive.
Before reading and after incorporating the substrate into the mixture, this is equally crucial.
