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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 (It does not recognize NAD+/NADH). There is no need to purify NADP/NADPH from sample mix. The enzyme cycling reaction significantly increases detection sensitivity. Results can be quantified using plate reader at OD450nm.
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If you would like to use a fluorometric reading, please refer to NADP/NADPH Assay Kit (Fluorometric) (ab176724).
|NADP Cycling Buffer||WM||1 x 15ml|
|NADP Cycling Enzyme Mix||Green||1 x 0.2ml|
|NADP/NADPH Extraction Buffer||NM||1 x 50ml|
|NADPH Developer||Purple||1 vial|
|NADPH Standard (MW:833.36)||Yellow||1 x 166.7µg|
|Stop Solution||Red||1 x 1.2ml|
NAPDH/NADP+ ratio was determined using ab65349 in stable HSPB1- knockdown U87 cells.
Standard curve with background signal subtracted (duplicates; +/- SD).
Total NADP and NADPH (tNADP) or NADPH alone measured in RAW cell lysates (duplicates +/- SD).
Measurement of NADPt and NADPH in rat liver lysate (20 μg). Assays were performed using the kit protocol.
Measurement of NADPt and NADPH in HeLa cell lysate (80 μg). Assays were performed using the kit protocol.
Typical standard curve for ab65349.
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