NADP/NADPH Assay Kit

NADP/NADPH Assay Kit (ab65349) provides a convenient tool for sensitive detection of the intracellular nucleotides: NADP, NADPH and their ratio. Assays of nicotinamide nucleotides are of continual interest in the studies of energy transforming and redox state of cells or tissue.

The enzymes in the system specifically recognize NADP/NADPH in an enzyme cycling reaction. The assay does not recognize NAD+/NADH. There is no need to purify NADP/NADPH from the sample mix. The enzyme cycling reaction significantly increases detection sensitivity. Results can be quantified using a plate reader at OD450nm.

NADP / NADPH assay protocol summary:
– extract samples from cells /  tissues with extraction buffer and deproteinize with spin column
– for NADPH measurement, heat samples to 60ºC for 30 min to decompose NAD⁺, cool on ice (this step not necessary for measurement of total NADP⁺/NADPH)
– add samples and standards to wells
– add reaction mix and incubate for 5 min at room temp to convert NADP to NADPH
– add NADPH developer and incubate for 1-4 hrs while reaction cycles
– analyze with microplate reader multiple times during the 1-4 hr incubation
– reaction can be stopped with stop solution
 

This product is manufactured by BioVision, an Abcam company and was previously called K347 NADP/NADPH Quantitation Colorimetric Kit. K347-100 is the same size as the 100 test size of ab65349.

If you would like to use a fluorometric reading, please refer to NADP/NADPH Assay Kit (Fluorometric) (ab176724).