Analysis of NAD+/NADH by Enzyme-labeled Instrument (CAT#: STEM-MB-1468-LGZ)

Determination of the total amount of NAD+ and NADH: ethanol under the action of enzyme to produce acetaldehyde, in this reaction process NAD+ is converted to NADH, NADH under the action of the electron coupled reagent to reduce WST-8 to orange substance, with a maximum absorption peak at about 450 nm. The amount of NAD+ and NADH in the reaction sample. The amount of NADH was determined separately: after the sample was extracted and heated in a 60°C water bath for 30 min, the NAD+ in the sample would decompose and only NADH would be retained. NADH reduces WST-8 to a yellow substance, and the amount of NADH in the sample can be determined separately. Determine the ratio of NAD+ and NAD+/NADH: The amount of NAD+ in the sample and the ratio of NAD+ to NADH can be obtained according to the total amount of NAD+ and NADH and the individual amount of NADH obtained by the detection in the first two steps. NADP+ and NADPH had no effect on the determination results due to the specificity of the enzyme reagents.

It was used to detect the content, ratio and total content of NAD+ and NADH in animal tissue and cell samples.

1. Prepare standard samples and experimental samples.
2. Add reaction reagents in order for reaction.
3. Measure the absorbance of each tube.
4. Make the mark curve and calculate the result.

• Sample Type: animal tissue and cell samples