Amplite® Fluorimetric NADP/NADPH Ratio Assay Kit *Red Fluorescence*

Amplite® Fluorimetric NADP/NADPH Ratio Assay Kit

Red Fluorescence

Amplite® Fluorimetric NADP/NADPH Ratio Assay Kit *Red Fluorescence* is a fluorimetric assay kit for quantification of NADP, NADPH, and their ratio in biological samples.

Detection Principle: Measures NADP/NADPH using an enzyme-cycling reaction.
Signal Advantage: Enzyme cycling enhances sensitivity, while red-range detection reduces biological sample interference.
Detection Chemistry: Uses an NADP/NADPH recycling enzyme mix with NADPH sensor buffer.
Workflow: Compatible with 96- or 384-well microplates and automation with no separation steps required.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm.

NADPH dose response was measured with Amplite® Fluorimetric NADP/NADPH Ratio Assay Kit in a 96-well solid black plate using a Gemini microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
15264	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® NADP/NADPH Ratio Assay Kit provides a convenient method for sensitive detection of NADP, NADPH and their ratio. The enzymes in the system specifically recognize NAD/NADH in an enzyme cycling reaction. There is no need to purify NADP/NADPH 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. This Amplite® Fluorimetric NADP/NADPH Assay Kit can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required.

Citations

View all 85 citations: Citation Explorer

Photothermal-Responsive Conjugated Polymer Nanoparticles Triggered Precise Augmentation of Microalgal Photosynthesis

Authors:

Zhang, Miaomiao and Gao, Chenying and Wang, Pengcheng and Li, Rui and Huang, Yiming and Lv, Fengting and Li, Cheng and Bai, Haotian and Zhang, Deqing and Wang, Shu

Journal:

ACS nano (2025)

Synergistic and stepwise treatment of resveratrol and catechol in Haematococcus pluvialis for the overproduction of biomass and astaxanthin

Authors:

Qiu, Jia-Fan and Yang, Yu-Cheng and Li, Ruo-Yu and Jiao, Yu-Hu and Mou, Jin-Hua and Yang, Wei-Dong and Lin, Carol Sze Ki and Li, Hong-Ye and Wang, Xiang

Journal:

Biotechnology for Biofuels and Bioproducts (2024): 80

The Function of HDAC6 Mediated Prx2 Acetylation in Neuronal Apoptosis Related Alzheimer's disease

Authors:

Zhang, Lijie and Hu, Jinxia and Zhang, Tao and Gao, Huimin and Wu, Xingrui and Zhang, Conghui and Zhang, Cheng and Chen, Hao and Yang, Minggang and Cao, Xichuan and others,

Solar-powered multi-organism symbiont mimic system for beyond natural synthesis of polypeptides from CO2 and N2

Authors:

Yu, Wen and Zeng, Yue and Wang, Zenghao and Xia, Shengpeng and Yang, Zhiwen and Chen, Weijian and Huang, Yiming and Lv, Fengting and Bai, Haotian and Wang, Shu

Journal:

Science Advances (2023): eadf6772

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

Authors:

Allie, Robert

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	576 ± 5 nm
Recommended plate	Clear bottom

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: NADP/NADPH Recycling Enzyme Mix	2 bottles (lyophilized powder)
Component B: NADPH Sensor Buffer	1 bottle (20 mL)
Component C: NADPH Standard	1 vial (167 µg)
Component D: NADPH Extraction Solution	1 bottle (10 mL)
Component E: NADP Extraction Solution	1 bottle (10 mL)
Component F: NADP/NADPH Control Solution	1 bottle (10 mL)
Component G: NADP/NADPH Lysis Buffer	1 bottle (10 mL)