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.

How the assay works
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.

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.

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

How other researchers are using 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}

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.