Best substrate of Alkaline Phosphatase?

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Adam B Shapiro

para-nitrophenylphosphate (PNPP) is a colorimetric substrate for alkaline phosphatase that produces a soluble product.

Nick Gee

pNPP is a great substrate and most people use it. A more sensitive fluorescent option is 4-methylumbelliferyl phosphate. If you need only a slightly stronger signal, but do not have a fluorescence plate reader, you can detect Pi that is liberated (instead of p-nitrophenol) using a malachite green-based reagent. This will give you about 4x the signal possible with p-nitrophenol.