Name: Amplite® Fluorimetric Total NAD and NADH Assay Kit *Red Fluorescence*
Brand: AAT Bioquest
SKU: 15257
Price: 355 USD
Availability: InStock

Amplite® Fluorimetric Total NAD and NADH Assay Kit

Red Fluorescence

Amplite® Fluorimetric Total NAD and NADH Assay Kit Red Fluorescence is a fluorimetric assay kit for detection of total NAD and NADH in a microplate format.

Detection Principle: Uses an enzyme-cycling reaction that specifically recognizes NAD/NADH.
Signal Advantage: Enzyme cycling enhances sensitivity, while red-range detection reduces biological sample interference.
Workflow: Measures NAD/NADH without requiring purification from the sample mixture.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm using solid black microplates.

NADH dose response was measured with Amplite® Total NAD and NADH Assay Kit in a solid black 96-well plate using a NOVOStar microplate reader (BMG Labtech). RFU at Ex/Em = 540/590 nm.

Protocol

SDS

Catalog	Size	Price	Quantity
15257	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. In addition, this assay has very low background since it is run in the red visible range that significantly reduces the interference from biological samples. The assay has demonstrated high sensitivity and low interference with 570 nm excitation 590 nm emission.

Citations

View all 66 citations: Citation Explorer

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

Authors:

Allie, Robert

NAMPT-dependent NAD+ salvage is crucial for the decision between apoptotic and necrotic cell death under oxidative stress

Authors:

Nishida, Takuto and Naguro, Isao and Ichijo, Hidenori

Journal:

Cell Death Discovery (2022): 1--11

B cell Sirt1 deacetylates histone and non-histone proteins for epigenetic modulation of AID expression and the antibody response

Authors:

Gan, Huoqun and Shen, Tian and Chupp, Daniel P and Taylor, Julia R and Sanchez, Helia N and Li, Xin and Xu, Zhenming and Zan, Hong and Casali, Paolo

Journal:

Science advances (2020): eaay2793

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)

Mitochondrial elongation-mediated glucose metabolism reprogramming is essential for tumour cell survival during energy stress

Authors:

Li, J and Huang, Q and Long, X and Guo, X and Sun, X and Jin, X and Li, Z and Ren, T and Yuan, P and Huang, X and others,

Journal:

Oncogene (2017): 4901--4912

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	540 nm
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

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Page updated on October 8, 2026

Component A: NAD/NADH Recycling Enzyme Mix	2 bottles (lyophilized powder)
Component B: NADH Sensor Buffer	1 bottle (20 mL)
Component C: NADH Standard (FW: 709)	1 vial (142 µg)