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Product Catalog |
NAD+/NADH Colorimetric Assay KitCode No: CY-1253
Target:
NAD+/NADH Background:
Nicotinamide adenine dinucleotide (NAD+) as well as nicotinamide adenine dinucleotide phosphate (NADP+) is an important cofactor found in cells. NADH is the reduced form of NAD+, and NAD+ is the oxidized form of NADH. It has been reported that NAD+ metabolism regulates important biological effects including life span. NAD+, through poly-ADP-ribosyl polymerase (PARP), mono-ADP- ribosyltransferase (ARTs) and recently characterized sirtuin enzymes, exerts potential biological effects. These enzymes modify proteins to regulate their function via ADP-ribosylation or deacetylation in the presence of NAD+. These enzymes are involved in several pathways including apoptosis, DNA repair, senescence and endocrine signaling, suggesting that either the enzymes could be an important therapeutical target for cancer, diabetes atherosclerosis and so on. The traditional NAD+/NADH and NADP+/NADPH assays are done by monitoring of NADH or NADPH absorption at 340 nm. This method suffers low sensitivity and high interference since the assay is done in the UV range that requires expensive quartz microplate. CycLex NAD+/NADH Colorimetric Assay Kit employs an enzyme cycling reaction, which significantly increases detection sensitivity, and provides a convenient method for sensitive detection of NAD+, NADH and their ratio. The measurement of NAD+ and NADH concentrations in most laboratories is possible if they are equipped with a microtiter plate reader. Considering that the use of fully automatic apparatus to monitor the absorbance has become widespread, NAD+ and NADH concentration measurement is now possible with the CycLex NAD+/NADH Colorimetric Assay Kit using the same equipment. This method of measurement should dramatically raise the efficiency of measuring NAD+ and NADH concentrations in mammalian cells and tissues.
Size:
100 assays
Product Type:
Kit
Storage Temp. (°C):
-70 References:
1. Ziegenhorn J, Senn M, Bucher T. (1976) Molar absorptivities of beta-NADH and beta-NADPH. Clin Chem. 22: 151. 2. Matsumura, H. and Miyachi S (1980) Cycling assay for nicotinamide adenine dinucleotides. Methods Enzymol. 69: 465-470. 3. Zerez CR, Lee SJ, Tanaka KR (1987) Spectrophotometric determination of oxidized and reduced pyridine nucleotides in erythrocytes using a single extraction procedure. Anal Biochem. 164: 367-73 4. Zhao, Z, Hu, X and Ross CW (1987). Comparison of Tissue Preparation Methods for Assay of Nicotinamide Coenzymes. Plant Physiol. 84: 987-988. 5 Umemura, K and Kimura, H (2005) Determination of oxidized and reduced nicotinamide adenine dinucleotide in cell monolayers using a single extraction procedure and a spectrophotometric assay. Anal Biochem. 338: 131-5. 6. Hasmann, M and Schemainda, I (2003) FK866, a Highly Specific Noncompetitive Inhibitor of Nicotinamide Phosphoribosyltransferase, Represents a Novel Mechanism for Induction of Tumor Cell Apoptosis. Cancer Research 63: 7436-7442, 2003 7.Vilcheze, C et al. (2005). Altered NADH/NAD+ Ratio Mediates Coresistance to Isoniazid and Ethionamide in Mycobacteria. Antimicrobial Agents and Chemotherapy. 49: 708-720. 8. Kimura N, Fukuwatari T, Sasaki R, Shibata K. (2006) Comparison of metabolic fates of nicotinamide, NAD+ and NADH administered orally and intraperitoneally; characterization of oral NADH. J Nutr Sci Vitaminol. (Tokyo) 52: 142. 9. O'Donnell JM, et al. (2004) Limited transfer of cytosolic NADH into mitochondria at high cardiac workload. Am J Physiol Heart Circ Physiol. 286: H2237. 10. Richard A. et al. (2008) Characterization of NAD Uptake in Mammalian Cells J. Biol. Chem. 283: 6367-6374. 11. Rongvaux, A et al. (2008) Nicotinamide Phosphoribosyl Transferase/Pre-B Cell Colony-Enhancing Factor/Visfatin Is Required for Lymphocyte Development and Cellular Resistance to Genotoxic Stress J. Immunol. 181: 4685-4695 12. Pogrebniak, A et al. (2006) Chemopotentiating effects of a novel NAD biosynthesis inhibitor, FK866, in combination with antineoplastic agents. Eur J Med Res. 11: 313-21.
Intended Use:
For Research use only. Not for use in diagnostic procedure. |
