Amplite® Colorimetric NAD/NADH Ratio Assay Kit

Amplite® Colorimetric NAD/NADH Ratio Assay Kit is a colorimetric assay kit for measuring NAD, NADH, and the NAD/NADH ratio in biological samples.

Colorimetric detection: Uses a chromogenic NADH probe with absorbance at approximately 460 nm.
High sensitivity: Detects as little as 3 µM NADH in a 100 µL assay volume.
Ratio measurement: Includes extraction and neutralization reagents for determining NAD and NADH separately.
Microplate compatible: Supports 96-well or 384-well formats.
Applications: Suitable for measuring cellular NAD/NADH redox status and metabolic activity.

Amplite® Colorimetric NAD/NADH Ratio Assay Kit is used to measure total NAD/NADH amount and in a white/clear 96-well microplate using a SpectraMax microplate reader (Molecular devices).

Protocol

SDS

Catalog	Size	Price	Quantity
15273	250 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. 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. Our Amplite® NAD/NADH Ratio Assay Kit provides a convenient method for sensitive detection of NAD, NADH and their ratio. The NADH probe is a chromogenic sensor that has its maximum absorbance at ~460 nm upon NADH reduction. The absorbance increase at ~460 nm is directly proportional to the concentration of NADH in the solution. The NADH probe can recognize NADH in an enzyme-free reaction, and the signal can be easily read by an absorbance microplate reader at ~460 nm. The Amplite® Colorimetric NADH Assay Kit provides a sensitive assay to detect as little as 3 µM NADH in a 100 µL assay volume. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format.

Citations

View all 90 citations: Citation Explorer

High salt exposure promotes tau hyperphosphorylation and mitochondrial reverse electron transport associated neurodegeneration in Drosophila

Authors:

Kasanin, Jacob and Lu, Benjamin G and Zheng, Xiwei and Khau, Kodi and Rimal, Suman

Journal:

Scientific Reports (2026)

Targeting reverse electron transport preserves mitochondrial function and prevents dopaminergic neurodegeneration in Parkinson’s disease

Authors:

Bhurtel, Sunil and Rimal, Suman and Li, Wen and Lu, Bingwei

Journal:

Molecular Neurodegeneration Advances (2026): 33

Reprogramming of the kynurenine pathway impairs NAD+ homeostasis and mediates doxorubicin-induced cardiotoxicity in mice

Authors:

Li, Danlei and Zhang, Yang and Kuang, Yuanyuan and Lin, Zhong and Wang, Ping and Jiang, Jianjun and Pi, Wenhu and Ma, Qilin

Journal:

Redox Biology (2025): 103957

Regulation of Energy Metabolism and Lipid Metabolites by IMP3 in Cervical Cancer

Authors:

Lei, Xiaojun and Yang, Wenyuan and Xu, Sen and Wu, Haigang

Journal:

Current Issues in Molecular Biology (2025): 1014

The role of metabolic enzyme acetylation in cardiac energy metabolism

Authors:

Ketema, Ezra Belay

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 12, 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: NAD Extraction Solution	1 bottle (10 mL)
Component E: Neutralization Solution	1 bottle (10 mL)
Component F: Extraction Control Solution	1 bottle (10 mL)
Component G: Lysis Buffer	1 bottle (10 mL)