Extraction and Quantitation of Nicotinamide Adenine Dinucleotide Redox Cofactors.

ABSTRACT:

Aims

Accurate analysis of dinucleotide redox cofactors nicotinamide adenine dinucleotide phosphate reduced (NADPH), nicotinamide adenine dinucleotide phosphate (NADP⁺), nicotinamide adenine dinucleotide reduced (NADH), and nicotinamide adenine dinucleotide (NAD⁺) from biological samples is important to understanding cellular redox homeostasis. In this study, we aimed to develop a simple protocol for quenching metabolism and extracting NADPH that avoids interconversion among the reduced forms and the oxidized forms.

Results

We compared seven different solvents for quenching and extraction of cultured mammalian cells and mouse tissues: a cold aqueous buffer commonly used in enzyme assays with and without detergent, hot aqueous buffer, and cold organic mix

SUBMITTER: Lu W 

PROVIDER: S-EPMC5737638 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature