PiPer™ Phosphate Assay Kit - FAQs

How does the PiPer Phosphate Assay kit work?

In the PiPer Phosphate Assay, maltose phosphorylase converts maltose (in the presence of Pi) to glucose1-phosphate and glucose. Then, glucose oxidase converts the glucose to gluconolactone and H₂O₂. Finally, with horseradish peroxidase (HRP) as a catalyst, the H₂O₂ reacts with the Amplex Red reagent (10-acetyl-3,7-dihydroxyphenoxazine) to generate resorufin, which has absorption and fluorescent emission maxima of 563/587 nm. The resulting increase in fluorescence or absorption is proportional to the amount of Pi in the sample. By fluorescence, the kit can be used to detect as little as 800 nM Pi.

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