Amplite® Fluorimetric Total NADP and NADPH Assay Kit *Red Fluorescence*

Amplite® Fluorimetric Total NADP and NADPH Assay Kit

Red Fluorescence

Amplite® Fluorimetric Total NADP and NADPH Assay Kit *Red Fluorescence* is a fluorimetric assay kit for quantification of total NADP and NADPH in a microplate format.

Detection Principle: Uses an enzyme-cycling reaction that specifically recognizes NADP/NADPH.
Signal Advantage: Enzyme cycling increases sensitivity, while red-range detection reduces interference from biological samples.
Workflow: Measures NADP/NADPH directly from sample mixtures without requiring purification.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm using solid black microplates; absorbance detection is also supported at 576 ± 5 nm.

NADPH dose response was measured with Amplite® Fluorimetric Total NADP and NADPH Assay Kit in a solid black 96-well plate using a NOVOStar microplate reader (BMG Labtech). RFU is Ex/EM = 540/590 nm.

Protocol

SDS

Catalog	Size	Price	Quantity
15259	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® NADP/NADPH Assay Kit provides a convenient method for sensitive detection of NADP and NADPH. The enzymes in the system specifically recognize NADP/NADPH 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 with 570 nm excitation 590 nm emission.

Citations

View all 61 citations: Citation Explorer

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)

Resveratrol attenuates excessive ethanol exposure induced insulin resistance in rats via improving NAD+/NADH ratio

Authors:

Luo, Gang and Huang, Bingqing and Qiu, Xiang and Xiao, Lin and Wang, Ning and Gao, Qin and Yang, Wei and Hao, Liping

Journal:

Molecular Nutrition & Food Research (2017)

Epigenetic regulation of Runx2 transcription and osteoblast differentiation by nicotinamide phosphoribosyltransferase

Authors:

Ling, Min and Huang, Peixin and Islam, Shamima and Heruth, Daniel P and Li, Xuanan and Zhang, Li Qin and Li, Ding-You and Hu, Zhaohui and Ye, Shui Qing

Journal:

Cell & Bioscience (2017): 27

MCU-dependent mitochondrial Ca2+ inhibits NAD+/SIRT3/SOD2 pathway to promote ROS production and metastasis of HCC cells

Authors:

Ren, T and Zhang, H and Wang, J and Zhu, J and Jin, M and Wu, Y and Guo, X and Ji, L and Huang, Q and Yang, H and others, undefined

Journal:

Oncogene (2017)

References

View all 40 references: Citation Explorer

NADPH recycling systems in oxidative stressed pea nodules: a key role for the NADP(+)-dependent isocitrate dehydrogenase

Authors:

Marino D, Gonzalez EM, Frendo P, Puppo A, Arrese-Igor C.

Journal:

Planta. (2006)

Glucose-6-phosphate dehydrogenase activity and NADPH/NADP+ ratio in liver and pancreas are dependent on the severity of hyperglycemia in rat

Authors:

Diaz-Flores M, Ibanez-Hern and ez MA, Galvan RE, Gutierrez M, Duran-Reyes G, Medina-Navarro R, Pascoe-Lira D, Ortega-Camarillo C, Vilar-Rojas C, Cruz M, Baiza-Gutman LA.

Journal:

Life Sci (2006): 2601

Titration of E. coli transhydrogenase domain III with bound NADP+ or NADPH studied by NMR reveals no pH-dependent conformational change in the physiological pH range

Authors:

Pedersen A, Johansson T, Rydstrom J, Goran Karlsson B.

Journal:

Biochim Biophys Acta (2005): 254

A novel NADPH:(bound) NADP+ reductase and NADH:(bound) NADP+ transhydrogenase function in bovine liver catalase

Authors:

Gaetani GF, Ferraris AM, Sanna P, Kirkman HN.

Journal:

Biochem J (2005): 763

Modulation of cooperativity in Mycobacterium tuberculosis NADPH-ferredoxin reductase: cation-and pH-induced alterations in native conformation and destabilization of the NADP+-binding domain

Authors:

Bhatt AN, Shukla N, Aliverti A, Zanetti G, Bhakuni V.

Journal:

Protein Sci (2005): 980

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)