NADP/NADPH Assay Kit (Fluorometric)
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Supplier Data
Functional Studies - NADP/NADPH Assay Kit (Fluorometric) (AB176724)
NADPH dose response was measured with ab176724 in a 96-well solid black plate using a microplate reader.
Product details
NADP/NADPH Assay Kit (Fluorometric) ab176724 provides a convenient method for sensitive detection of NADP, NADPH and their ratio.
The enzymes in the system specifically recognize NADP/NADPH in an enzyme recycling reaction that significantly increases detection sensitivity. In addition, this assay has very low background since it is run in the red visible range that considerably reduces the sample interference.
The NADP/NADPH assay can be performed in a 96-well or 384-well microtiter-plate format. The signal can be read by either a fluorescence microplate reader at Ex/Em = 530-570/590-600 nm (maximum Ex/Em = 540/590 nm) or an absorbance microplate reader at ~576 nm.
NADP/ NADPH assay protocol summary:
- add samples and standards to wells
- add extraction solutions and incubate for 15 min
- add reaction mix and incubate for 30 min - 2 hr
- analyze with microplate reader
Other Notes
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. NAD 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 used as reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis.
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Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NADPH facilitates reductive biosynthesis and maintains cellular redox status. It acts as a major electron donor in various anabolic reactions including lipid and nucleic acid synthesis. NADPH also contributes to the detoxification of reactive oxygen species and serves in the process of oxidative phosphorylation. Its cofactor activity associates it predominantly with enzymatic complexes such as the cytochrome P450 system.
Pathways
NADPH is fundamental in both the pentose phosphate pathway and the Calvin cycle. In the pentose phosphate pathway it provides reducing power important for fatty acid and cholesterol biosynthesis. In photosynthetic organisms NADPH functions in the Calvin cycle facilitating carbon fixation. The enzyme glucose-6-phosphate dehydrogenase related to NADPH production is integral in NADPH recycling.
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Publications (19)
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Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association 191:114845 PubMed38945390
2024
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The Journal of clinical investigation 134: PubMed38869951
2024
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Redox biology 74:103234 PubMed38861834
2024
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The EMBO journal 43:2636-2660 PubMed38778156
2024
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The Journal of biological chemistry 299:105186 PubMed37611829
2023
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eLife 12: PubMed37449973
2023
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The Journal of biological chemistry 299:104920 PubMed37321450
2023
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Proceedings of the National Academy of Sciences of the United States of America 119:e2204527119 PubMed35858325
2022
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Journal of experimental botany 72:5534-5552 PubMed33974689
2021
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Frontiers in microbiology 12:617050 PubMed33584619
2021
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