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
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Extract samples from cells / tissues with extraction buffer and
deproteinize with spin column
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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)
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Add samples and standards to wells
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Add reaction mix and incubate for 5 min at room temp to convert NAD to
NADH
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Add NADH developer and incubate for 1-4 hrs while reaction cycles
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Analyze with microplate reader multiple times during the 1-4 hr incubation
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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:
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Human: primary blood mononuclear cells1, epithelial ovarian
cancer cells2, Jurkat cells3
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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.
