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Why p-nitrophenolphosphate → p-nitrophenol is an ideal reaction for this laboratory assay?

Sagot :

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.

What is p-nitrophenolphosphate ?

  • 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.

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