Continuous PRPP-S Assay Kit

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Continuous PRPP-S Assay Kit

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Cat.No. : Kit-0871
Product Overview : PRPP-S Assay Kit is designed to measure PRPP (α-D-5-phosphoribosyl-1-pyrophosphate) content in samples. This enzymatic assay is based on a coupled reaction involving Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) and Inosine Monophosphate Dehydrogenase (IMPDH).
The principle of the assay is based on the coupling of the following enzymatic reactions:
(1) In the presence of ATP and P-ribose, PRPP-Synthetase enzyme catalyzes the formation of PRPP.
(2) In the presence of Hypoxanthine (Hx), PRPP is converted to IMP by Hypoxanthine-guanine phosphoribosyltransferase (HGPRT).
(3) IMP is immediately oxidized by a highly active IMPDH in the presence of NAD with simultaneous formation of NADH2 directly monitored spectrophotometrically at 340nm.
The assay is developed for measuring PRPP-S activity in vitro or in cell lysates.
For maximal accuracy, the assays with cell lysates are run with and without P-ribose 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 : 24 analysis

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