NAD/NADH Assay Kit (Colorimetric)

NAD/NADH Assay Kit (Colorimetric) ab65348 provides a convenient and sensitive tool to quantify NAD⁺ and NADH, and measure their ratio, in samples from mammals and other species.

The NAD cycling enzyme mix in the kit specifically acts on NADH/NAD in an enzyme cycling reaction which significantly increases sensitivity and specificity. There is no requirement to purify NADH/NAD from samples.

The levels of both NADt (total NAD⁺ and NADH) and NADH can be easily measured; the level of NAD⁺ can be easily calculated by subtracting NADH from NADt. The assay is read by absorbance at 450 nm.

NAD / NADH assay protocol summary:
- extract samples from cells / tissues with extraction buffer and deproteinize with spin column
- for NADH measurement, heat samples to 60°C for 30 min to decompose NAD⁺, cool on ice (this step not necessary for measurement of total NAD⁺/NADH)
- add samples and standards to wells
- add reaction mix and incubate for 5 min at room temp to convert NAD to NADH
- add NADH developer and incubate for 1-4 hrs while reaction cycles
- analyze with microplate reader multiple times during the 1-4 hr incubation
- reaction can be stopped with stop solution.

This assay specifically detects NAD and NADH, but not NADP nor NADPH. If you would like to use a fluorometric reading, please refer to NAD/NADH Assay Kit (Fluorometric) (NAD/NADH Assay Kit (Fluorometric) ab176723). NAD/NADH Assay kit NAD/NADH Assay Kit II (colorimetric) ab221821 uses an alternative assay method that relies on purification of NAD and NADH from samples and may be more sensitive in some samples. Review our to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader. **How other researchers have used NAD/NADH Assay Kit ab65348** This NAD/NADH assay kit has been used in publications in a variety of sample types, including: - Human: primary blood mononuclear cells1, epithelial ovarian cancer cells2, Jurkat cells3 - Mouse: cell culture lysates4, cardiomyocyte cell culture lysates5, liver6, liver and muscle7, primary hepatocyte cell cultures8, aorta tissue9 - Rat: brain tissue10 - Locust: thoraic muscle11 Bacteria: Z mobilis12, E coli13 References: 1 - Castro-Marrero J et al 2015; 2 - Zhu J et al 2019, Xia H et al 2015; 3 - Miller TW et al 2015; 4 - Mekala NK et al 2019, Ling S et al 2017; 5 - Zhang D et al 2019; 6 - Shao D et al 2017, Mukherji A et al 2015, Yu JH et al 2016; 7 - Karandrea S et al 2017; 8 - Traboulsi H et al 2014; 9 - Liu Y et al 2016; 10 - Rao G et al 2016; 11 - Ding D et al 2018; 12 - Wu B et al 2019; 13 - Long YM et al 2017