Amplite® Colorimetric Total NAD and NADH Assay Kit *Enhanced Sensitivity*

Amplite® Colorimetric Total NAD and NADH Assay Kit

Enhanced Sensitivity

Amplite® Colorimetric Total NAD and NADH Assay Kit *Enhanced Sensitivity* is a colorimetric assay kit for quantifying total NAD and NADH in biological samples.

Detection Principle: Uses an enzyme cycling reaction for NAD/NADH detection.
Sensitivity: Provides higher sensitivity than Kit #15258.
Specificity: Enzymes specifically recognize NAD/NADH.
Workflow: No NAD/NADH purification is required.
Instrument Compatibility: Absorbance detection at 460 nm.

NADH dose response was measured with the Amplite® Colorimetric Total NAD and NADH Assay Kit *Enhanced Sensitivity* in a 96-well white/clear bottom plate using a SpectraMax microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
15275	400 Tests	Price

Overview

Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+, and NAD+ is the oxidized form of NADH. It forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. NADP is used in anabolic biological reactions, such as fatty acid and nucleic acid synthesis, which require NADPH as a reducing agent. In chloroplasts, NADP is an oxidizing agent important in the preliminary reactions of photosynthesis. The NADPH produced by photosynthesis is then used as reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis. The traditional NAD/NADH and NADP/NADPH assays are done by monitoring of NADH or NADPH absorption at 340 nm. This method suffers low sensitivity and high interference since the assay is done in the UV range that requires expensive quartz microplate. This Amplite® NAD/NADH Assay Kit provides a convenient method for sensitive detection of NAD and NADH. The enzymes in the system specifically recognize NAD/NADH in an enzyme cycling reaction. There is no need to purify NAD/NADH from sample mix. The enzyme cycling reaction significantly increases detection sensitivity. Compared to Kit #15258, this kit has higher sensitivity.

Citations

View all 61 citations: Citation Explorer

NAD+ Synthetase is Required for Free-living and Symbiotic Nitrogen Fixation in the Actinobacterium Frankia casuarinae

Authors:

Kucho, Ken-ichi and Asukai, Koya and Van Nguyen, Thanh

Journal:

Microbes and Environments (2023): ME22093

NAD (H)-loaded nanoparticles for efficient sepsis therapy via modulating immune and vascular homeostasis

Authors:

Ye, Mingzhou and Zhao, Yi and Wang, Yuyuan and Xie, Ruosen and Tong, Yao and Sauer, John-Demian and Gong, Shaoqin

Journal:

Nature Nanotechnology (2022): 1--11

The Linkage of Yeast Metabolites, Produced Under Hyperosmotic Stress, to Cellular Cofactor Systems During Icewine Fermentation.

Authors:

Allie, Robert

Enhanced 1, 3-propanediol production in Klebsiella pneumoniae by a combined strategy of strengthening the TCA cycle and weakening the glucose effect

Authors:

Lu, Xinyao and Ren, Shunli and Lu, Jingzheng and Zong, Hong and Song, Jian and Zhuge, Bin

Journal:

Journal of applied microbiology (2018)

Cytosolic Redox Status of Wine Yeast (Saccharomyces Cerevisiae) under Hyperosmotic Stress during Icewine Fermentation

Authors:

Yang, Fei and Heit, Caitlin and Inglis, Debra L

Journal:

Fermentation (2017): 61

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	460 nm
Recommended plate	Clear bottom

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Page updated on October 8, 2026

Component A: NAD/NADH Recycling Enzyme Mix	2 bottles (lyophilized powder)
Component B-I: NADH Probe	1 bottle (4 mL)
Component B-II: NADH Probe Buffer	1 bottle (16 mL)
Component C: NADH Standard (FW: 709)	1 vial (142 µg)
Component D: Lysis Buffer	1 bottle (10 mL)