EnzyChrom™ NADP/NADPH Assay Kit
Protocol SDS  Printed Protocols and SDSs are not provided with kits.

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
Hu, Li, et al (2019). Melatonin decreases M1 polarization via attenuating mitochondrial oxidative damage depending on UCP2 pathway in prorenin-treated microglia. PloS one 14.2: e0212138. Assay: NADP/NADPH in rat microglial cells.

You, Sung-Hwan, et al (2019). Rapid and sensitive detection of NADPH via mBFP-mediated enhancement of its fluorescence. PloS one 14.2: e0212061. Assay: NADP/NADPH in bacteria cells.

Jiang, L-B., et al (2018). TIGAR mediates the inhibitory role of hypoxia on ROS production and apoptosis in rat nucleus pulposus cells. Osteoarthritis and cartilage 26.1: 138-148. Assay: NADP/NADPH in rat cells.

Khadka, Dipendra, et al (2018). Augmentation of NAD+ levels by enzymatic action of NAD (P) H quinone oxidoreductase 1 attenuates adriamycin-induced cardiac dysfunction in mice. Journal of molecular and cellular cardiology 124: 45-57. Assay: NADP/NADPH in mice tissues.

Kim, Jae-Eung, et al (2018). Rerouting of NADPH synthetic pathways for increased protopanaxadiol production in Saccharomyces cerevisiae. Scientific reports 8.1: 15820. Assay: NADP/NADPH in yeast cells.

Kwak, Min-Kyu, MyungHee Ku, and Sa-Ouk Kang (2018). Inducible NAD (H)-linked methylglyoxal oxidoreductase regulates cellular methylglyoxal and pyruvate through enhanced activities of alcohol dehydrogenase and methylglyoxal-oxidizing enzymes in glutathione-depleted Candida albicans. Biochimica et Biophysica Acta (BBA)-General Subjects 1862.1: 18-39. Assay: NADP/NADPH in C. albicans (fungi) cells.

Upadhyay, Rashmi, et al (2018). Host Directed Therapy for Chronic Tuberculosis via Intrapulmonary Delivery of Aerosolized Peptide Inhibitors Targeting the IL-10-STAT3 Pathway. Scientific reports 8.1: 16610. Assay: NADP/NADPH in mice tissues.

Wang, Weixuan, et al (2018). ROS-Mediated 15-Hydroxyprostaglandin Dehydrogenase Degradation via Cysteine Oxidation Promotes NAD+-Mediated Epithelial-Mesenchymal Transition. Cell chemical biology 25.3: 255-261. Assay: NADP/NADPH in human cells.

Fu, Ling, et al (2017). Systematic and quantitative assessment of hydrogen peroxide reactivity with cysteines across human proteomes. Molecular & Cellular Proteomics 16.10: 1815-1828. Assay: NADP/NADPH in human cells.

Ou, Weijun, et al (2017). Low-density lipoprotein docosahexaenoic acid nanoparticles induce ferroptotic cell death in hepatocellular carcinoma. Free Radical Biology and Medicine 112: 597-607. Assay: NADP/NADPH in human cells.

Shin, YoungHo, et al (2017). Cytochrome c peroxidase regulates intracellular reactive oxygen species and methylglyoxal via enzyme activities of erythroascorbate peroxidase and glutathione-related enzymes in Candida albicans. The international journal of biochemistry & cell biology 92: 183-201. Assay: NADP/NADPH in C. albicans (fungi) cells.

Hatori, Yuta, et al (2016). Neuronal differentiation is associated with a redox-regulated increase of copper flow to the secretory pathway. Nature communications 7: 10640. Assay: NADP/NADPH in human cells.

Zhou, Jun-Hao, et al (2016). TIGAR contributes to ischemic tolerance induced by cerebral preconditioning through scavenging of reactive oxygen species and inhibition of apoptosis. Scientific reports 6: 27096. Assay: NADP/NADPH in mice cells.

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.

Lee, W.H., 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 Escherichia coli bacteria.

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.

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. 92(5):929-37. Assay: NADP/NADPH in Escherichia coli bacteria.

Tang, G.J., 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 lung epithelial cells.

Brown, S.M., 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 Cryptococcus neoformans yeast.

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.

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 panceatic cancer cells.

Lee, H.C., 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 Escherichia coli bacteria.

Lui, V.W., 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 cancer cell lines.

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 cells.

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 mice muscle tissue.

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.

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.

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 Synechocystis sp. cyanobacteria.

Lee, H.C., 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 Escherichia coli bacteria.

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

Tseng, H.C., 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 Escherichia coli bacteria.

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.

To find more recent publications, please click here.
Please inquire or request assay service or call 1-510-782-9988 x 2.
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EnzyChrom™ NADP/NADPH Assay Kit
Catalog No: ECNP-100
Price: $369    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: On Ice
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: -20°C

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