Continuous Phosphatase Assay Kit

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Continuous Phosphatase Assay Kit

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Cat.No. : Kit-0881
Product Overview : Phosphatase Assay Kit provides a simple one-step spectrophotometric method for the quantitation of inorganic phosphate (Pi) in solution, including Pi released from enzymatic reactions. This kit enables continuous assay of reactions that generate Pi such as those catalyzed by ATPases and GTPases.
The release of inorganic phosphate is measured through coupling to PNP-XDH enzymatic system. In the presence of inorganic phosphate, PNP (purine nucleoside phosphorylase) catalyzes inosine phosphorolysis leading to the formation of hypoxanthine and P-ribose. XDH enzyme catalyzes irreversible oxidation of hypoxanthine to uric acid with simultaneous reduction of NAD to NADH2, measurable by absorbance at 340nm. The formation of NADH2 is continuously monitored spectrophotometrically at 340nm. The assay is linear in 5-300μM range.
For maximal accuracy, the assays with cell lysates are run with and without inosine in parallel. The absorbance rate observed in the absence of inosine is used as blank and is subtracted from the absorbance rate measured in its presence.
Storage : Once dissolved, the reagents provided in the kit are not stable and should be stored on ice and used the day of preparation.
Size : 1 plate (96 assays)

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