| Field | Specification |
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| Product Type | |
| Sample Type(s) | Cell or tissue extracts |
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Overview
For sensitive determination of NAD and NADH and evaluation of drug effects on NAD/NADH metabolism. The assay uses OD565nm for signal readout. Compatible sample input includes Cell or tissue extracts. Typical stated assay timing is 15 min.
Key elements and design rationale
- Readout format: OD565nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Cell or tissue extracts, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.05 µM for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. The detection limit of 0.05 µM and linearity up to 10 µM NAD+/NADH in 96-well plate assay.
Additional feature notes highlight Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and 15 min at room temperature; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of nad/nadh within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
Pyridine nucleotides play an important role in metabolism and, thus, there is continual interest in monitoring their concentration levels. Quantitative determination of NAD+/NADH has applications in research pertaining to energy transformation and the redox state of cells or tissue. Simple, direct, and automation-ready procedures for measuring NAD+/NADH concentration are very desirable. BioAssay Systems EnzyChrom™ NAD+/NADH assay kit is based on a lactate dehydrogenase cycling reaction, in which the formed NADH reduces a formazan (MTT) reagent. The intensity of the reduced product color, measured at 565 nm, is proportional to the NAD+/NADH concentration in the sample. This assay is highly specific for NAD+/NADH and with minimal interference (<1%) by NADP+/NADPH. Our assay is a convenient method to measure NAD, NADH, and their ratio.
Detection method
Colorimetric (OD 565 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.05 µM.
Procedures and timing
Stated procedure or timing information: 15 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify nad/nadh in cell or tissue extracts by OD565 nm readout.
- Compare treatment or phenotype groups using matched cell or tissue extracts handling.
- Monitor time-course or pre/post changes in cell or tissue extracts across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
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 laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Relevance of nadh dehydrogenase and alternative two-enzyme systems for growth of corynebacterium glutamicum with glucose, lactate, and acetate
Maeda, T., et al (2021). Relevance of nadh dehydrogenase and alternative two-enzyme systems for growth of corynebacterium glutamicum with glucose, lactate, and acetate. Frontiers in Bioengineering and Biotechnology, 8. Assay: NAD/NADH in C. glutamicum cells.
D4F prophylaxis enables redox and energy homeostasis while preventing inflammation during hypoxia exposure
Paul, S., et al (2021). D4F prophylaxis enables redox and energy homeostasis while preventing inflammation during hypoxia exposure. Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie, 133, 111083. Assay: NAD/NADH in male Sprague Dawley rats lung tissue.
Nicotinamide promotes differentiation of pancreatic endocrine progenitors from human pluripotent stem cells through poly (ADP-ribose) polymerase inhibition
Woodford, C., et al (2020). Nicotinamide promotes differentiation of pancreatic endocrine progenitors from human pluripotent stem cells through poly (ADP-ribose) polymerase inhibition. BioRxiv, 2020.04.21.052951. Assay: NAD/NADH in mouse cells.
Metabolic engineering of 1,2-propanediol production from cellobiose using beta-glucosidase-expressing E
Nonaka, D., et al (2021). Metabolic engineering of 1,2-propanediol production from cellobiose using beta-glucosidase-expressing E. coli. Bioresource Technology, 329, 124858. Assay: NAD/NADH in E. coli cells.
Phenformin inhibits hedgehog-dependent tumor growth through a complex i-independent redox/corepressor module
Di Magno, L., et al (2020). Phenformin inhibits hedgehog-dependent tumor growth through a complex i-independent redox/corepressor module. Cell Reports, 30(6), 1735-1752.e7. Assay: NAD/NADH in Math1-CRE;Ptch1 tissue.
A reduced form of nicotinamide riboside defines a new path for NAD+ biosynthesis and acts as an orally bioavailable NAD+ precursor
Giroud-Gerbetant, J., et al (2019). A reduced form of nicotinamide riboside defines a new path for NAD+ biosynthesis and acts as an orally bioavailable NAD+ precursor. Molecular Metabolism, 30, 192-202. Assay: NAD/NADH in Mice tissue and cells.
Subcellular NAMPT-mediated NAD+ salvage pathways and their roles in bioenergetics and neuronal protection after ischemic injury
Wang, X., et al (2019). Subcellular NAMPT-mediated NAD+ salvage pathways and their roles in bioenergetics and neuronal protection after ischemic injury. Journal of Neurochemistry, 151(6), 732-748. Assay: NAD/NADH in C57BL/6J mice neurons.
Li, R., et al (2020, August 7). Emodin alleviates hydrogen peroxide-induced inflammation and oxidative stress via mitoch
Li, R., et al (2020, August 7). Emodin alleviates hydrogen peroxide-induced inflammation and oxidative stress via mitochondrial dysfunction by inhibiting the pi3k/mtor/gsk3β pathway in neuroblastoma sh-sy5y cells [Research Article]. BioMed Research International. Assay: NAD/NADH in SH-SY5Y cells.
Xu, K., et al (2020, December 30). Vgll4 protects against oxidized-ldl-induced endothelial cell dysfunction and inflamma
Xu, K., et al (2020, December 30). Vgll4 protects against oxidized-ldl-induced endothelial cell dysfunction and inflammation by activating hippo-yap/tead1 signaling pathway [Research Article]. Mediators of Inflammation. Assay: NAD/NADH in Ox-LDL-induced human umbilical vein endothelial cells.
Metabolic engineering of shikimic acid-producing corynebacterium glutamicum from glucose and cellobiose retaining its phosphotransferase system function and pyruvate kinase activities
Sato, N., et al (2020). Metabolic engineering of shikimic acid-producing corynebacterium glutamicum from glucose and cellobiose retaining its phosphotransferase system function and pyruvate kinase activities. Frontiers in Bioengineering and Biotechnology, 8. Assay: NAD/NADH in C. glutamicum cells cells.
Synergistic effect and mechanism of plumbagin with gentamicin against carbapenem-resistant klebsiella pneumoniae
Chen, X., et al (2020). Synergistic effect and mechanism of plumbagin with gentamicin against carbapenem-resistant klebsiella pneumoniae. Infection and Drug Resistance, 13, 2751-2759. Assay: NAD/NADH in Klebsiella pneumoniae cells.
Emodin inhibits zinc-induced neurotoxicity in neuroblastoma SH-SY5Y cells
Liu, W., et al (2019). Emodin inhibits zinc-induced neurotoxicity in neuroblastoma SH-SY5Y cells. Bioscience Reports, 39(BSR20182378). Assay: NAD/NADH in Human neuroblastoma SH-SY5Y cells.
Effects of nadh availability on 3-phenyllactic acid production by lactobacillus plantarum expressing formate dehydrogenase
Li, M., et al (2019). Effects of nadh availability on 3-phenyllactic acid production by lactobacillus plantarum expressing formate dehydrogenase. Current Microbiology, 76(6), 706-712. Assay: NAD/NADH in Lactobacillus plantarum cells.
Application of a pyruvate-producing escherichia coli strain lafcpcpt-accbc-acee: A case study for d-lactate production
Wada, K., et al (2020). Application of a pyruvate-producing escherichia coli strain lafcpcpt-accbc-acee: A case study for d-lactate production. Fermentation, 6(3), 70. Assay: NAD/NADH in E. coli HIT-DH5α cells.
Deletion of CD38 and supplementation of NAD + attenuate axon degeneration in a mouse facial nerve axotomy model
Takaso, Y., et al (2020). Deletion of CD38 and supplementation of NAD + attenuate axon degeneration in a mouse facial nerve axotomy model. Scientific Reports, 10(1), 17795. Assay: NAD/NADH in mouse cells.
Both gain and loss of Nampt function promote pressure overload-induced heart failure
Byun, J., et al (2019). Both gain and loss of Nampt function promote pressure overload-induced heart failure. American Journal of Physiology-Heart and Circulatory Physiology, 317(4), H711-H725. Assay: NAD/NADH in mouse cells.
Sato, N., et al (2020a). Metabolic engineering of shikimic acid-producing corynebacterium glutamicum from glucose and ce
Sato, N., et al (2020a). Metabolic engineering of shikimic acid-producing corynebacterium glutamicum from glucose and cellobiose retaining its phosphotransferase system function and pyruvate kinase activities. Frontiers in Bioengineering and Biotechnology, 8. Assay: NAD/NADH in C. glutamicum cells.
Characterization of balofloxacin-stressed proteomics and identification of balofloxacin-binding proteins pre-peptidase and integration host factor in Edwardsiella tarda
Wen, Q., et al (2019). Characterization of balofloxacin-stressed proteomics and identification of balofloxacin-binding proteins pre-peptidase and integration host factor in Edwardsiella tarda. Journal of Proteomics, 205, 103413. Assay: NAD/NADH in E. tarda cells.
Deletion of Topoisomerase 1 in excitatory neurons causes genomic instability and early onset neurodegeneration
Fragola, G., et al (2020). Deletion of Topoisomerase 1 in excitatory neurons causes genomic instability and early onset neurodegeneration. Nature Communications, 11(1), 1962. Assay: NAD/NADH in Mouse cortices.
L-lysine potentiates aminoglycosides against Acinetobacter baumannii via regulation of proton motive force and antibiotics uptake
Deng, W., et al (2020). L-lysine potentiates aminoglycosides against Acinetobacter baumannii via regulation of proton motive force and antibiotics uptake. Emerging Microbes & Infections, 9(1), 639-650. Assay: NAD/NADH in A. baumannii cells.
A bacterial cell factory converting glucose into scyllo -inositol, a therapeutic agent for Alzheimer?s disease
Michon, C., et al (2020). A bacterial cell factory converting glucose into scyllo -inositol, a therapeutic agent for Alzheimer?s disease. Communications Biology, 3(1), 1-7. Assay: NAD/NADH in B. subtilis cells.
Metabolic regulation of endothelial SK channels and human coronary microvascular function
Liu, Y., et al (2020). Metabolic regulation of endothelial SK channels and human coronary microvascular function. International Journal of Cardiology, 312, 1-9. Assay: NAD/NADH in Human myocardial cells.
Pharmacological restoration of autophagy reduces hypertension-related stroke occurrence
Forte, M., et al (2020). Pharmacological restoration of autophagy reduces hypertension-related stroke occurrence. Autophagy, 16(8), 1468-1481. Assay: NAD/NADH in Human A10 cells.
Essentiality of fatty acid synthase in the 2D to anchorage-independent growth transition in transforming cells
Bueno, M. J., et al (2019). Essentiality of fatty acid synthase in the 2D to anchorage-independent growth transition in transforming cells. Nature Communications, 10(1), 5011. Assay: NAD/NADH in nude mice cells.
Nicotinamide pathway-dependent sirt1 activation restores calcium homeostasis to achieve neuroprotection in spinocerebellar ataxia type 7
Stoyas, C. A., et al (2020). Nicotinamide pathway-dependent sirt1 activation restores calcium homeostasis to achieve neuroprotection in spinocerebellar ataxia type 7. Neuron, 105(4), 630-644.e9. Assay: NAD/NADH in Mouse cerebellar granule neurons.
Walker, M. A., et al (2021a). Acetylation of muscle creatine kinase negatively impacts high-energy phosphotransfer in he
Walker, M. A., et al (2021a). Acetylation of muscle creatine kinase negatively impacts high-energy phosphotransfer in heart failure. JCI Insight, 6(3). Assay: NAD/NADH in Mouse cardiac tissue.
Inhibition of glucose assimilation in Auxenochlorella protothecoides by light
Xiao, Y., et al (2020). Inhibition of glucose assimilation in Auxenochlorella protothecoides by light. Biotechnology for Biofuels, 13(1), 146. Assay: NAD/NADH in Auxenochlorella protothecoides cells.
Bioluminescent-based imaging and quantification of glucose uptake in vivo
Maric, T., et al (2019). Bioluminescent-based imaging and quantification of glucose uptake in vivo. Nature Methods, 16(6), 526-532. Assay: NAD/NADH in 4T1-luc cells.
Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits
Kilfoil, P. J., et al (2019). Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits. Journal of Molecular and Cellular Cardiology, 137, 93-106. Assay: NAD/NADH in mice heart tissue.
The effect of NAMPT deletion in projection neurons on the function and structure of neuromuscular junction (Nmj) in mice
Lundt, S., et al (2020). The effect of NAMPT deletion in projection neurons on the function and structure of neuromuscular junction (Nmj) in mice. Scientific Reports, 10(1), 99. Assay: NAD/NADH in mice muscle tissue.
Increasing ascomycin yield in streptomyces hygroscopicus var
Wang, P., et al (2020). Increasing ascomycin yield in streptomyces hygroscopicus var. Ascomyceticus by using polyhydroxybutyrate as an intracellular carbon supply station. Assay: NAD/NADH in S. hygroscopicus var. ascomyceticus FS35 hyphae.
Khattab, S. M. R., & Watanabe, T. (2021a). Metabolic engineering of Saccharomyces cerevisiae for efficient conversions o
Khattab, S. M. R., & Watanabe, T. (2021a). Metabolic engineering of Saccharomyces cerevisiae for efficient conversions of glycerol to ethanol. BioRxiv, 2021.01.04.425180. Assay: NAD/NADH in E. coli cells.
Integrated Regulation of Cardiac Fatty Acid and Glucose Oxidation
Altamimi, T. R. (2018). Integrated Regulation of Cardiac Fatty Acid and Glucose Oxidation. Assay: NAD/NADH in mice heart tissue.
STAT3-RXR-Nrf2 activates systemic redox and energy homeostasis upon steep decline in pO2 gradient
Paul, S., Gangwar, A., Bhargava, K., & Ahmad, Y. (2018). STAT3-RXR-Nrf2 activates systemic redox and energy homeostasis upon steep decline in pO2 gradient. Redox biology, 14, 423-438. Assay: NAD/NADH in sprague dewley rats tissue.
ROS-Mediated 15-Hydroxyprostaglandin Dehydrogenase Degradation via Cysteine Oxidation Promotes NAD+-Mediated Epithelial-Mesenchymal Transition
Wang, W., Hu, Y., Wang, X., Wang, Q., & Deng, H. (2018). ROS-Mediated 15-Hydroxyprostaglandin Dehydrogenase Degradation via Cysteine Oxidation Promotes NAD+-Mediated Epithelial-Mesenchymal Transition. Cell chemical biology, 25(3), 255-261. Assay: NAD/NADH in mice tissue.
Reconstruction of metabolic module with improved promoter strength increases the productivity of 2-phenylethanol in Saccharomyces cerevisiae
Wang, Z., Jiang, M., Guo, X., Liu, Z., & He, X. (2018). Reconstruction of metabolic module with improved promoter strength increases the productivity of 2-phenylethanol in Saccharomyces cerevisiae. Microbial cell factories, 17(1), 60. Assay: NAD/NADH in yeast cells.
The aldehyde group of gossypol induces mitochondrial apoptosis via ROS-SIRT1-p53-PUMA pathway in male germline stem cell
He, X., Wu, C., Cui, Y., Zhu, H., Gao, Z., Li, B. & Zhao, B. (2017). The aldehyde group of gossypol induces mitochondrial apoptosis via ROS-SIRT1-p53-PUMA pathway in male germline stem cell. Oncotarget, 8(59), 100128. Assay: NAD/NADH in cotton cells.
The transcriptional regulator of the chaperone response HSF1 controls hepatic bioenergetics and protein homeostasis
Qiao, A., Jin, X., Pang, J., Moskophidis, D., & Mivechi, N. F. (2017). The transcriptional regulator of the chaperone response HSF1 controls hepatic bioenergetics and protein homeostasis. J Cell Biol, 216(3), 723-741. Assay: NAD/NADH in mice liver tissue.
Efficient production of acetoin in Saccharomyces cerevisiae by disruption of 2, 3-butanediol dehydrogenase and expression of NADH oxidase
Bae, S. J., Kim, S., & Hahn, J. S. (2016). Efficient production of acetoin in Saccharomyces cerevisiae by disruption of 2, 3-butanediol dehydrogenase and expression of NADH oxidase. Scientific reports, 6, 27667. Assay: NAD/NADH in yeast cells.
PARP-1 inhibition increases mitochondrial metabolism through SIRT1 activation
Bai P et al (2011). PARP-1 inhibition increases mitochondrial metabolism through SIRT1 activation. Cell Metab. 13(4):461-8. Assay: NAD/NADH in mouse cells.
Improvement of coenzyme Q(10) production by increasing the NADH/NAD(+) ratio in Agrobacterium tumefaciens
Koo BS et al (2010). Improvement of coenzyme Q(10) production by increasing the NADH/NAD(+) ratio in Agrobacterium tumefaciens. Biosci Biotechnol Biochem.74(4):895-8. Assay: NAD/NADH in yeast Agrobacterium tumefaciens.
Depletion of GSH in glial cells induces neurotoxicity: relevance to aging and degenerative neurological diseases
Lee M et al (2010). Depletion of GSH in glial cells induces neurotoxicity: relevance to aging and degenerative neurological diseases. FASEB J. 24(7):2533-45. Assay: NAD/NADH in human cell.
Nicotinamide phosphoribosyltransferase regulates cell survival through NAD+ synthesis in cardiac myocytes
Hsu CP et al (2009). Nicotinamide phosphoribosyltransferase regulates cell survival through NAD+ synthesis in cardiac myocytes. Circ Res. 105(5):481-91. Assay: NAD/NADH in mouse heart cardiac myocytes.
Metabolic engineering of Escherichia coli for enhanced production of (R)- and (S)-3-hydroxybutyrate
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: NAD/NADH in bacteria E.coli.
Small-molecule inhibition of 6-phosphofructo-2-kinase activity suppresses glycolytic flux and tumor growth
Clem B,et al (2008). Small-molecule inhibition of 6-phosphofructo-2-kinase activity suppresses glycolytic flux and tumor growth. Mol Cancer Ther. 7(1):110-20. Assay: NAD/NADH in human epithelial cell.
A genome wide analysis of the response to uncapped telomeres in budding yeast reveals a novel role for the NAD+ biosynthetic gene BNA2 in chromosome end protection
Greenall A et al (2008). A genome wide analysis of the response to uncapped telomeres in budding yeast reveals a novel role for the NAD+ biosynthetic gene BNA2 in chromosome end protection. Genome Biol. 9(10):R146. Assay: NAD/NADH in yeast cell.
Dihydrolipoamide dehydrogenase mutation alters the NADH sensitivity of pyruvate dehydrogenase complex of Escherichia coli K-12
Kim Y, et al (2008). Dihydrolipoamide dehydrogenase mutation alters the NADH sensitivity of pyruvate dehydrogenase complex of Escherichia coli K-12. J Bacteriol. 190(11):3851-8. Assay: NAD/NADH in bacterial E. coli.
Anticancer agent CHS-828 inhibits cellular synthesis of NAD
Olesen UH, et al (2008). Anticancer agent CHS-828 inhibits cellular synthesis of NAD. Biochem Biophys Res Commun. 367(4):799-804. Assay: NAD/NADH in human cell.
Visfatin: a new player in mesangial cell physiology and diabetic nephropathy
Song HK, et al (2008). Visfatin: a new player in mesangial cell physiology and diabetic nephropathy. Am J Physiol Renal Physiol. 295(5):F1485-94. Assay: NAD/NADH in human mesangial cells.
Targeting aspartate aminotransferase in breast cancer
Thornburg JM et al (2008). Targeting aspartate aminotransferase in breast cancer. Breast Cancer Res. 10(5):R84. Assay: NAD/NADH in human breast adenocacinoma cell.
