LS-K299-100
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sILSBio LifeSpan BioSciences, Inc. Ethanol Assay Kit (Colorimetric) LS-K299-100 (100 Tests) • Store at -20°C Introduction Alcoholic drinks are among the daily consumed beverages. Studies have shown heavy alcohol consumption may lead to various forms of liver diseases and to increased mortality rates. Quantitative determination of alcohol (ethanol, C2H5OH) has applications in basic research, drug discovery, clinic studies and in the alcoholic industry. Simple, direct and automation-ready procedures for measuring ethanol concentration are very desirable. LSBio's ethanol assay kit is based on alcohol dehydrogenase catalyzed oxidation of ethanol, in which the formed NADH is coupled to the formazan (MTT) chromogen. The intensity of the product color, measured at 565 nm, is proportionate to the ethanol concentration in the sample. Key Features • Sensitive and accurate. Detection limit 0.0008 vol % (140 pM or 8 ppm), linearity up to 0.1% ethanol in 96-well plate assay. • Convenient. The procedure involves adding working reagent, incubating for 30 min and stopping reaction. No 37°C heater is needed. • High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Applications • Direct Assays: ethanol in serum, plasma, urine and saliva samples. • Pharmacology: effects of drugs on alcohol metabolism. Materials Not Supplied Pipetting (multi-channel) devices. Clear flat-bottom 96-well plates (e.g.Corning Costar) and plate reader. Storage The kit is shipped on ice. Store all kit components at -20 °C. FOR RESEARCH USE ONLY! Not for use in humans. LifeSpan BioSciences, Inc. • 2401 Fourth Avenue, Suite 900, Seattle, WA 98121 www.LSBio.com • (206) 464-1554 • TechnicalSupport@LSBio.com

sILSBio LifeSpan BioSciences, Inc. Ethanol Assay Kit (Colorimetric) LS-K299-100 (100 Tests) • Store at -20°C Assay Procedure Reagent Preparation: Reconstitute Enzyme A by adding 120 pL Enzyme Buffer to the Enzyme A tube. Make sure Enzyme A is fully dissolved by pipetting up and down. Store reconstituted Enzyme A at -20°C and use within 1 month. Assay Procedure 1. Calibration Curve. Prepare 0.1% alcohol Premix by mixing 25 pL 1% Standard and 225 pL distilled water. Dilute standard as follows. Transfer 10 pL standards into wells of a clear flat-bottom 96-well plate. Samples: add 10 pL sample...

sILSBio LifeSpan BioSciences, Inc. LifeSpan BioSciences, Inc. • 2401 Fourth Avenue, Suite 900, Seattle, WA 98121 www.LSBio.com • (206) 464-1554 • TechnicalSupport@LSBio.com