Fluorescent NADP/NADPH Detection Kit

A non-fluorescent detection reagent is reduced in the presence of NADPH to produce its fluorescent analog and NADP. NADP is further converted to NADPH via an enzyme-coupled reaction. The kit specifically detects NADP/NADPH and not NAD/NADH.



SKU: NADPH100-2

Size: 100 Tests
Price:
Sale price$591

Nicotinamide adenine dinucleotide phosphate (NADP+) is used in anabolic reactions, such as lipid and nucleic acid synthesis, which require NADPH as a reducing agent. NADPH is the reduced form of NADP+, and NADP+ is the oxidized form of NADPH. In cells, NADPH plays the role of a carrier of reducing power and is primarily involved in maintaining optimal redox metabolism. A simplified assay for the measurement of NAD and NADP is critical to understanding the roles of these pyridine nucleotides in normal and abnormal cells.

NADPH is produced in the oxidative phase of the pentose phosphate pathway in cells, a multifunctional pathway whose primary purpose is to generate reducing power in the form of NADPH. NADPH is a cofactor for enzymes that synthesize energy-rich molecules and provide the reducing equivalents for the oxidation-reduction involved in protecting the cell from the toxicity of reactive oxygen species (ROS) and NADPH oxidase-dependent ROS generation. Both NAD and NADP have been shown to influence hemoglobin affinity for oxygen in erythrocytes. In plant cells, NADPH is used as the reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis.

Our Fluorescent NADP/NADPH Detection Kit provides a reliable, sensitive fluorometric assay for the quantification of NADP, NADPH and their ratio in biological samples. The kit can be used with mammalian cells and tissues, and bacterial, fungal and plant cells.

The kit utilizes a non-fluorescent detection reagent, which is reduced in the presence of NADPH to produce its fluorescent analog and NADP. NADP is further converted to NADPH via an enzyme-coupled reaction. The enzymes in the reactions specifically react with NADP/NADPH and not with NAD/NADH.

Fluorescence plate reader
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NADPH100-2: 100 Tests
  • Part # 6022: Enzyme Mix, 170μL x 1 (2-8C)
  • Part # 3050: NADP/NADPH Lysis Solution, 12.5mL x 1 (2-8C)
  • Part # 3057 (Green label): NADP Extraction Buffer, 22 mL x 1 (2-8C)
  • Part # 3058 (Magenta label): NADPH Extraction Buffer, 22 mL x 1 (2-8C)
  • Part # 3049: Reaction Buffer, 11 mL x 1 (2-8C)
  • Part # 3052: Standard curve diluent, 15 mL x 1 (2-8C)
  • Part # 4019: NADPH Detection reagent, 110 μL x 1 (-20C)
  • Part # 3048: 3X Substrate Mix, 6mL x 1 (-20C)
  • Part # 7014: NADPH Standard, 3 vials dried (-20C)

  • NADPH100-3: 500 Tests
  • Part # 6022: Enzyme Mix, 170μL x 5 (2-8C)
  • Part # 3050: NADP/NADPH Lysis Solution, 12.5mL x 5 (2-8C)
  • Part # 3057 (Green label): NADP Extraction Buffer, 22 mL x 5 (2-8C)
  • Part # 3058 (Magenta label): NADPH Extraction Buffer, 22 mL x 5 (2-8C)
  • Part # 3049: Reaction Buffer, 11 mL x 5 (2-8C)
  • Part # 3052: Standard curve diluent, 15 mL x 5 (2-8C)
  • Part # 4019: NADPH Detection reagent, 110 μL x 5 (-20C)
  • Part # 3048: 3X Substrate Mix, 6mL x 5 (-20C)
  • Part # 7014: NADPH Standard, 3 vials dried (-20C)
  • Product Specific References

    PMID Publication
    33168978Kobayashi, H., et al. 2021. Chemical reversal of abnormalities in cells carrying mitochondrial DNA mutations. Nat Chem Biol, 335-343.
    31234468Dougan, J., et al. 2019. Proteomics-Metabolomics Combined Approach Identifies Peroxidasin as a Protector against Metabolic and Oxidative Stress in Prostate Cancer. Int J Mol Sci, 3046.
    29847794Petit, L., et al. 2018. Aerobic Glycolysis Is Essential for Normal Rod Function and Controls Secondary Cone Death in Retinitis Pigmentosa. Cell Rep, 2629-2642.
    28285191Wu, H., et al. 2017. Polychlorinated biphenyls-153 induces metabolic dysfunction through activation of ROS/NF-κB signaling via downregulation of HNF1b. Redox Biol, 300-310.
    28598329Chinchore, Y., et al. 2017. Glycolytic reliance promotes anabolism in photoreceptors. Elife, e25946.
    28566553Ghanegolmohammadi, F., et al. 2017. Systematic analysis of Ca2+ homeostasis in Saccharomyces cerevisiae based on chemical-genetic interaction profiles. Mol Biol Cell, 3415-3427.
    25882048Itsumi, M., et al. 2015. Idh1 protects murine hepatocytes from endotoxin-induced oxidative stress by regulating the intracellular NADP+/NADPH ratio. Cell Death Differ, 1837-45.
    25798619Venkatesh, A., et al. 2015. Activated mTORC1 promotes long-term cone survival in retinitis pigmentosa mice. J Clin Invest, 1446-58.
    25070093Wang, J.H., et al. 2014. Sigma S-Dependent Antioxidant Defense Protects Stationary-Phase Escherichia coli against the Bactericidal Antibiotic Gentamicin. Antimicrob Agents Chemother, 5964-75.
    23734909Fridman, A., et al. 2013. Cell cycle regulation of purine synthesis by phosphoribosyl pyrophosphate and inorganic phosphate. Biochem J, 91-9.
    22925884Sasaki, M., et al. 2012. D-2-hydroxyglutarate produced by mutant IDH1 perturbs collagen maturation and basement membrane function . Genes Dev, 2038-49.
    22389629Nishida, K., et al. 2012. Induction of biogenic magnetization and redox control by a component of the target of rapamycin complex 1 signaling pathway. PLoS Biol, e1001269.

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