Ethanol Assay Kit

Antigen: Ethanol

Application: Biochemical Assay (BCA)

Catalog no.: ABIN411656

Additional Information

Sample Type: Cell and Tissue Culture Supernatant, Urine, Plasma, Serum, Biological Fluids

Components: Ethanol Assay Buffer, Ethanol Probe (in DMSO,anhydrous), Ethanol Enzyme Mix (lyophilized), Ethanol Standard (MW:46.07, 17.15N)

Description: Alcohol (ethanol C 2 H 5 OH) is among the most widely consumed drinks. Low doses of alcohol may help circulation while heavy alcohol consumption may lead to various forms of disease. Quantitative determination of alcohol finds applications in basic research, drug discovery, clinical studies and fermentation industry processes. The newly improved Ethanol Assay Kit is more sensitive and stable. The kit can accurately quantify ethanol concentration in a variety of biological samples such as serum, plasma, other body fluids, foods, beverages and growth media. In the assay, alcohol oxidase oxidizes ethanol to generate H 2 O 2 which reacts with our probe to generate color (λ max = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit detects 0.1-10 ppm alcohol (~10-800 nM). Contents Components Volume Cap color Part Number Ethanol Assay Buffer Ethanol Probe (in DMSO,anhydrous) Ethanol Enzyme Mix (lyophilized) Ethanol Standard (MW:46.07, 17.15N) 25 ml 200 µl 1 vial 0.5 ml WM Red Green Yellow K620-100-1 K620-100-2A K620-100-4 K620-100-5 III. Reagent Preparation: Ethanol Probe : Ready to use as supplied. Warm to room temperature prior to use. Store at –20 o C, avoid contamination with water, protect from light. Use within two months. Ethanol Enzyme Mix: Dissolve the Ethanol Enzyme Mix in 220 μ l Ethanol Assay Buffer prior to use. Store at 4 o C. Use within two months. IV. Ethanol

Application Details

Protocol: NOTE: THE ASSAY IS VERY SENSITIVE TO ALCOHOL VAPOR. EXTREME CARE SHOULD BE TAKEN TO ENSURE THAT NO ALCOHOL VAPORS (ETHANOL, METHANOL, PROPANOL) ARE IN THE LABORATORY AIR WHERE THE ASSAY IS TO BE PERFORMED. LABORATORIES WHERE HPLC EQUIPMENT AND SOLVENTS ARE STANDING OR WHERE ALCOHOL IS USED TO WIPE DOWN LABORATORY BENCHES OR EQUIPMENT ARE INAPPROPRIATE LOCATIONS TO PERFORM THIS ASSAY. 1. Standard Curve Preparations: For the colorimetric assay, add 50 μ l of pure ethanol standard (4 o C, ice cold) to 821.4 μ l Ethanol Assay Buffer, mix well. Then take 10 μ l of the dilution into 990 μ l assay buffer to generate 10 nmol/ μ l of ethanol standard. Take 100 μ l of the dilution into 900 μ l assay buffer to generate 1mM (1nmol/ µl). Add 0, 2, 4, 6, 8, 10 μ l to a series of wells in a 96 well plate and adjust the volume of each to 50 μ l with Assay Buffer to generate 0, 2, 4, 6, 8, 10 nmol/well ethanol Standard. Note: The fluorometric assay is 10-fold more sensitive than the colorimetric assay. For fluorometric measurement, dilute the ethanol standard to 0.1mM by adding 100 μ l of the ethanol standard to 900 μ l of Reaction Buffer, mix well. Then add 0, 2, 4, 6, 8, 10 μ l to a series of wells and adjust volume of each to 50 μ l with Assay Buffer to generate 0, 0.2, 0.4, 0.6, 0.8, 1.0 nmol/well ethanol Standard. 2. Sample Preparation: Samples can be diluted directly in Assay Buffer and tested. Biological samples such as serum (containing ~ 0.01-0.16% w/v) should be diluted 1:10-1:100 and volumes in the range of 10-30µl used. For beverages which contain 100X more alcohol, correspondingly greater dilutions should be used. We suggest making several dilutions of your sample so that the sample reading is within the standard curve range. When too much ethanol is in the reaction, the reaction develops from pink color to purple to brown, then becomes colorless. 3. Reaction Mix Preparation: Mix enough reagent for the number of assays performed: For each well, prepare a total 50 μ l Reaction Mix containing: 46 μ l Ethanol Assay Buffer 2 μ l Ethanol Probe* 2 μ l Ethanol Enzyme Mix *

Storage: -20°C

Expiry Date: 12 months

Restrictions: For Research Use only

Publications

Lu, Wu, Wang et al.: "Chronic alcohol-induced liver injury and oxidant stress are decreased in cytochrome P4502E1 knockout mice and restored in humanized cytochrome P4502E1 knock-in mice." in: Free radical biology & medicine, Vol. 49, Issue 9, pp. 1406-16, 2010 (PubMed).

Product: Inokuchi, Tsukamoto, Park et al.: "Toll-Like Receptor 4 Mediates Alcohol-Induced Steatohepatitis Through Bone Marrow-Derived and Endogenous Liver Cells in Mice." in: Alcoholism, clinical and experimental research, 2011 (PubMed).