Wednesday, August 16, 2017

NADH Analysis Just Got a Lot Easier!

The next generation 800™ TS Absorbance Reader was recently introduced in a previous blog posting. The many enhancements to both hardware and software were designed to simplify workflows for many of the most commonly performed assays. Assays such as ELISA, UV-absorption and colorimetric absorbance are performed in a variety of laboratory settings ranging from basic research in academic labs to clinical laboratory testing.

A long studied enzyme family found in a diverse range of organisms includes a group of alcohol dehydrogenase enzymes (ADH) which are responsible for interconversion between alcohols and aldehydes or ketones. The enzymatic catalysis of alcohol requires the reduction of the cofactor nicotinamide adenine dinucleotide (NAD+) to the reduced form NADH or the reverse chemical process for the formation of alcohol.

NADH

The spectral properties of the reduced form NADH, show an absorption peak at 340 nm providing a simple method to monitor progression of a reaction.  The enzymatic activity of ADH from S. cerevisiae was determined for the two alcohols, ethanol and 2-propanol, by obtaining kinetic absorbance measurements at 340 nm. Reaction velocities were calculated using Gen5™ Data Analysis software; Michaelis-Menten and Lineweaver Burk plots were generated to determine K­m and Vmax for each reaction.


These and similar experiments are made simple by the capabilities of the new, easy-to-use 800 TS. The 800 TS brings excellent performance, high quality and value to many laboratories. It's no wonder that BioTek continues to be the most trusted and well-known name in microplate readers and other life science instrumentation.


By: BioTek Instruments, Peter J. Brescia Jr., MSc, MBA

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