Abcam, ab176724, NADP/NADPH Assay Kit (Fluorometric)

Size: 250Test
NADP/NADPH Assay Kit (Fluorometric) ab176724 provides a convenient method for sensitive detection of NADP, NADPH and their ratio.
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Cell culture extracts, Tissue Extracts,
Assay type:Quantitative,
Assay time:1h,
Assay Platform: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.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
NADP/NADPH also known as Nicotinamide Adenine Dinucleotide Phosphate and its reduced form plays an essential role in cellular metabolism. It is a coenzyme with a molecular mass of around 743.4 g/mol. NADP/NADPH is ubiquitously expressed in cells due to its integral role in metabolic processes. It is found in high concentrations within the cytoplasm and chloroplasts where it functions in various biochemical reactions.
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.
Impaired NADPH function links to conditions like chronic granulomatous disease and diabetes. Chronic granulomatous disease occurs due to defects in the NADPH oxidase complex impairing the body's ability to combat pathogens effectively. Diabetes has associations with oxidative stress influenced by imbalance in NADPH levels. In these diseases NADPH's interaction with proteins like superoxide dismutase affects oxidative damage management and overall immune response.