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Is Amplite® Fluorimetric NAD/NADH Ratio Assay Kit *Red Fluorescence* pH sensitive? Do I have to prepare my samples in a certain pH buffer? Will pH influence my fluorescence readings?
Posted August 22, 2018
nicotinamide adenine dinucleotidenicotinamide adenine dinucleotide phosphateNADNADHNADH AssayNAD/NADH Ratio
Answer
pH is a factor during either the NAD or NADH extraction steps. The NADH extraction step (and corresponding buffer, Component D) is basic (pH >7) while the NAD extraction step (and corresponding buffer, Component E) is acidic (pH < 7). Additionally, as with most fluorescent probes, our NADH sensor is pH sensitive. Readings should be conducted under neutral conditions (pH≈7).