NAD/NADH Fluorometric Detection Kit

Common Name :

NAD/NADH

Cat.No. :

Kit-0614

Product Overview :

This NAD/NADH Fluorometric Detection Kit provides a convenient method for sensitive detection of NAD, NADH and their ratio. The enzymes in the system specifically recognize NAD/NADH in an enzyme cycling 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 interference from biological samples. There is no need to purify NAD/NADH from sample mix. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format. Its signal can be easily 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. This kit provides NAD and NADH extraction buffer, and cell lysis buffer for your convenience. It has been frequently used for determining NAD/NADH from cell lysates.

Description :

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. The traditional NAD/NADH and NADP/NADPH assays are done by monitoring the changes in 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 microplates.

Storage :

Keep in freezer Avoid exposure to light.

Size :

250 assays

Kit Components :

Component A: NAD/NADH Recycling Enzyme Mixture, 2 bottles (lyophilized powder)
Component B: NADH Sensor Buffer, 1 bottle (20 mL)
Component C: NADH Standard, 1 vial (142 µg)
Component D: NADH Extraction Solution, 1 bottle (10 mL)
Component E: NAD Extraction Solution, 1 bottle (10 mL)
Component F: NAD/NADH Control Solution, 1 bottle (10 mL)
Component G: NAD/NADH Lysis Buffer, 1 bottle (10 mL)

Detection method :

Fluorescence