SensoLyte NADP/NADPH Assay Ultra-sensitive & fluorimetric


AnaSpec, EGT Group, the provider of ultra-sensitive fluorimetric SensoLyte line of assay kits is pleased to announce the release of the SensoLyte Fluorimetric NADP/NADPH Assay Kit. This assay is a sensitive fluorimetric assay that detects NADP and NADPH without interfering with NAD/NADH. NADP is converted to NADPH in an enzyme-cycling reaction. The kit contains a fluorogenic reagent, producing fluorescence when reacted with NADPH. The resulting red fluorescence can be monitored at excitation /emission= 560/590 nm, with the intensity of fluorescence produced being proportional to NADP/NADPH concentration. This assay detects as low as 1.5 nM of the analyte.

Pyridine nucleotides are involved in a number of critical catabolic and anabolic reactions in living organisms. Nicotinamide adenine dinucleotide phosphate (NADP) is a natural coenzyme present in all animal and plant organisms. Cellular NADPH is important for tolerance to ROS and maintenance of cellular redox homeostasis.1-5 The reducing power of NADPH is a required cofactor for enzymes that are prooxidant such as nitric oxide synthase and NADPH oxidase.6 Furthermore the NADP+/NADPH ratio has been found to change in the erythrocytes of subjects affected by hemolytic disorder.7, 8

Product

Catalog #

SensoLyte NADP/NADPH Assay Kit *Fluorimetric*

72206

In addition to the fluorimetric kit, AnaSpec also provides a convenient colorimetric kit. This kit also detects NADP and NADPH without interfering with NAD/NADH. The SensoLyte Colorimetric NADP/NADPH Assay Kit utilizes an enzyme cycling reaction to produce a blue colored product, formazan, which can be monitored at 565 nm using a microplate reader. The intensity of the color produced is proportional to NADP/NADPH concentration.

Product

Catalog #

SensoLyte NADP/NADPH Assay Kit *Colorimetric*

72205



Fig 1. NADPH and NADH were serially diluted in assay buffer containing Enzyme Cycling Mix and Detection Reagent, and fluorescence was recorded at Ex/Em=570 /590 nm. (Flexstation 384II, Molecular Devices).



Fig 2. NADP and NAD were serially diluted in assay buffer containing Enzyme Cycling Mix and Detection Reagent, and fluorescence was recorded at Ex/Em=570/590 nm. (Flexstation 384II, Molecular Devices).


References:

1. Holmgren, A. J. Biol. Chem. 264, 13963 (1989).
2. Grant, CM. et al. Mol. Microbiol 21, 171 (1996a).
3. Muller, EG. et al. Mol. Biol.Cell 7, 1805 (1996).
4. Ng, CH. et al. Free Radic. Biol. Med. 44, 1131 (2008).
5. Temple, MD. et al. Trends Cell Biol. 15, 319 (2005).
6. Stamler, JS. et al. Science 258, 1898 (1992).
7. Kirkman, HN. et al. J. Clin. Invest. 55, 875 (1975).
8. Magnani, M. Acta Haematol. 75, 211 (1986).