How to calculate kinetic parameters with NADPH coupled reactions?

28 February 2014 1 8K Report

Do we have to subtract or add the blank reading (which lacks enzyme), from the reading with protein? I am not able to understand the calculations. Please help me out.

Shiv K. Sah-Teli

Hi, Yes you have to subtract the blank ( everything except enzyme) measurement from your enzyme catalyzed measurement. You might be monitoring the disappearance of NADPH, if so use extinction coefficient of 6,220 m–1 cm–1 at 340 nm for NADPH.

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