EnzyChrom™ NADP/NADPH Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of NADP and NADPH and evaluation of drug effects on their metabolism.

Key Features
Sensitive and accurate. Detection limit 0.1 μM, linearity up to 10 μM NADP+/NADPH in 96-well plate assay.

Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and 30 min at room temperature. No 37°C heater is required.

High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method
OD565nm

Samples
Cell, tissue extracts etc

Species
All

Procedure
30 min

Size
100 tests

Detection Limit
0.1 μM

Shelf Life
6 months

More Details
Pyridine nucleotides play an important role in metabolism and, thus, there is continual interest in monitoring their concentration levels. Quantitative determination of NADP+/NADPH has applications in research pertaining to energy transformation and redox state of cells or tissue. Simple, direct and automation-ready procedures for measuring NADP+/NADPH concentration are very desirable. BioAssay Systems’ EnzyChrom™ NADP+/NADPH assay kit is based on a glucose dehydrogenase cycling reaction, in which the formed NADPH reduces a formazan (MTT) reagent. The intensity of the reduced product color, measured at 565 nm, is proportionate to the NADP+/NADPH concentration in the sample. This assay is highly specific for NADP+/NADPH and is not interfered by NAD+/NADH. Our assay is a convenient method to measure NADP, NADPH and their ratio.

1. When tissue is used, should it be freshly obtained? Or is -80°C storage ok?



If direct sample processing is not possible, we recommend snap freezing tissue samples in liquid nitrogen and keeping them either in liquid nitrogen or at -80°C until further processing.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Korn, A et al (2009). Ferredoxin:NADP+ oxidoreductase association with phycocyanin modulates its properties. J Biol Chem 284(46):31789-97. Assay: NADP/NADPH in Cyanobacteria Synechocystis sp. (Pubmed).

2. Du, J et al (2010). Mechanisms of ascorbate-induced cytotoxicity in pancreatic cancer. Clin Cancer Res 16(2):509-20. Assay: NADP/NADPH in Human MIA PaCa-2 panceatic cancer cells (Pubmed).

3. Summermatter, S et al (2010). Peroxisome proliferator-activated receptor {gamma} coactivator 1{alpha} (PGC-1{alpha}) promotes skeletal muscle lipid refueling in vivo by activating de novo lipogenesis and the pentose phosphate pathway. J Biol Chem 285(43):32793-800. Assay: NADP/NADPH in human, mouse tissue (Pubmed).

4. Domain, F et al (2007). Transketolase A, an enzyme in central metabolism, derepresses the marRAB multiple antibiotic resistance operon of Escherichia coli by interaction with MarR. Mol Microbiol 66(2):383-94. Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

5. Guo, S et al (2009). Specific inhibition of hypoxia inducible factor 1 exaggerates cell injury induced by in vitro ischemia through deteriorating cellular redox environment. J Neurochem 108(5):1309-21. Assay: NADP/NADPH in Human neuroblastoma cells (Pubmed).

6. Tseng, HC et al (2009). Metabolic engineering of Escherichia coli for enhanced production of (R)- and (S)-3-hydroxybutyrate. Appl Environ Microbiol 75(10):3137-45. Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

7. Liu, M et al (2009). Cardiac Na+ current regulation by pyridine nucleotides. Circ Res 105(8):737-45. Assay: NADP/NADPH in Human HEK human embryonic kidney cells (Pubmed).

8. Lee, HC et al (2009). Thymidine production by overexpressing NAD+ kinase in an Escherichia coli recombinant strain. Biotechnol Lett 31(12):1929-36. Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

9. Lui, VW et al (2010). An RNA-directed nucleoside anti-metabolite, 1-(3-C-ethynyl-beta-d-ribo-pentofuranosyl)cytosine (ECyd), elicits antitumor effect via TP53-induced Glycolysis and Apoptosis Regulator (TIGAR) downregulation. Biochem Pharmacol 79(12):1772-80. Assay: NADP/NADPH in Human Nasopharyngeal cancer cell lines (Pubmed).

10. Brown, SM et al (2010). Isocitrate dehydrogenase is important for nitrosative stress resistance in Cryptococcus neoformans, but oxidative stress resistance is not dependent on glucose-6-phosphate dehydrogenase. Eukaryot Cell 9(6):971-80. Assay: NADP/NADPH in Yeast Cryptococcus neoformans (Pubmed).

11. Lee, HC et al (2010). High NADPH/NADP+ ratio improves thymidine production by a metabolically engineered Escherichia coli strain. J Biotechnol 149(1-2):24-32. Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

12. Osada-Oka, M et al (2010). Glucose is necessary for stabilization of hypoxia-inducible factor-1alpha under hypoxia: contribution of the pentose phosphate pathway to this stabilization. FEBS Lett 584(14):3073-9. Assay: NADP/NADPH in Human Hepatocellular carcinoma cell line Hep3B (Pubmed).

13. Tang, GJ et al (2011). Novel role of AMP-activated protein kinase signaling in cigarette smoke induction of IL-8 in human lung epithelial cells and lung inflammation in mice. Free Radic Biol Med 50(11):1492-502. Assay: NADP/NADPH in Human Bronchial epithelial cells (Pubmed).

14. Lee, WH et al (2011). Enhanced production of GDP-L: -fucose by overexpression of NADPH regenerator in recombinant Escherichia coli. Appl Microbiol Biotechnol 91(4):967-76. Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

15. Siedler, S et al (2011). Increased NADPH availability in Escherichia coli: improvement of the product per glucose ratio in reductive whole-cell biotransformation. Appl Microbiol Biotechnol . Assay: NADP/NADPH in Bacteria E.coli (Pubmed).

16. Ding, X et al (2009). Enhanced HtrA2/Omi expression in oxidative injury to retinal pigment epithelial cells and murine models of neurodegeneration. Invest Ophthalmol Vis Sci 50(10):4957-66. Assay: NADP/NADPH in Mouse Serum (Pubmed).

17. Cao, X et al (2010). The effects of quercetin in cultured human RPE cells under oxidative stress and in Ccl2/Cx3cr1 double deficient mice. Exp Eye Res 91(1):15-25. Assay: NADP/NADPH in Mouse Serum (Pubmed).

18. Csak, T et al (2011). Deficiency in myeloid differentiation factor-2 and toll-like receptor 4 expression attenuates nonalcoholic steatohepatitis and fibrosis in mice. Am J Physiol Gastrointest Liver Physiol 300(3):G433-41. Assay: NADP/NADPH in Mouse liver tissue extract (Pubmed).

19. Matsunami, T et al (2011). Enhancement of reactive oxygen species and induction of apoptosis in streptozotocin-induced diabetic rats under hyperbaric oxygen exposure. Int J Clin Exp Pathol 4(3):255-66. Assay: NADP/NADPH in Rat Pancreas (Pubmed).

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EnzyChrom™ NADP/NADPH Assay Kit
Catalog No: ECNP-100
Price: $349    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

Shipping: On Ice
Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: -20°C



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