EnzyChrom™ NAD/NADH Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For sensitive determination of NAD and NADH and evaluation of drug effects on NAD/NADH metabolism.

Key Features
Sensitive and accurate. Detection limit of 0.05 μM and linearity up to 10 μM NAD+/NADH in 96-well plate assay.

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.

Method
OD565nm

Samples
Cell or tissue extracts

Species
All

Procedure
15 min

Size
100 tests

Detection Limit
0.05 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 NAD+/NADH has applications in research pertaining to energy transformation and 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.

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. 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 (Pubmed).

2. 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 (Pubmed).

3. 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 Bacteria E.coli (Pubmed).

4. 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 (Pubmed).

5. 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 (Pubmed).

6. 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 (Pubmed).

7. 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 (Pubmed).

8. 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 (Pubmed).

9. 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 (Pubmed).

10. 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 (Pubmed).

11. 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 (Pubmed).

To find more recent publications, please click here.
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EnzyChrom™ NAD/NADH Assay Kit
Catalog No: E2ND-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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