Ethanol Colorimetric/Fluorometric Assay Kit

Catalog #: K620 | abID: ab65343

Product Details

abID ab65343
Cat # +Size K620-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit detects 0.1-10 ppm alcohol (~10-800 nM).
Features & Benefits • Simple procedure; takes ~40 minutes - 1 hour
• Fast and convenient
• Kit contains the necessary reagents for accurate measurement of ethanol in biological samples
Kit Components • Ethanol Assay Buffer
• Ethanol Probe (in DMSO,anhydrous)
• Ethanol Enzyme Mix (lyophilized)
• Ethanol Standard (MW:46.07, 17.15N)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Alcohol (ethanol C₂H₅OH) 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. BioVision’s Ethanol Assay Kit provides a simple, rapid, and sensitive method for accurate quantification of ethanol concentration in a variety of biological samples such as serum, plasma, other body fluids, foods, beverages and growth media. Alcohol oxidase oxidizes ethanol to generate H₂O₂ which reacts with our probe to generate color (λmax= 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit detects 0.1-10 ppm alcohol (~10-800 nM).

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Will this kit detect Isopropanol?
No, the kit is specific for ethanol as the enzyme used cannot detect isopropanol.
Can the initial dilution of the standard prep curve for the colorimetric assay (50 uL pure ethanol standard to 808.7 uL Ethanol Assay Buffer) be aliquoted and frozen for future use?
It is not recommended to freeze the pure Ethanol standard because some ethanol might vaporize changing the concentration of the solution during thawing. The pure ethanol standard should be prepared fresh ideally.
A flat std. curve is obtained but the values are very high. Zero ethanol std. curve point shows high values
This indicates there has been methanol/ethanol/propanol absorption into the buffer or other components. This is a classic diagnostic feature when the zero std. shows high values and due to saturation, the std. curve is flat. Extreme care should be taken to perform this assay in places where alcohol has not been used on the benches or there is no chance of contamination in the air. The fluorometric assay being at least 10 times more sensitive, can show larger interference due to contamination. Even incubating samples with ethanol next to blank might sometimes result in cross-contamination.
What kind of anti-coagulants should be used for blood/plasma/serum sample type used for this kit?
Heparin can be used with this kit.
Why is there pink color in the reaction mix? Isn't color development only possible when there is sample/ethanol?
The probe is colorless to begin with. But it is very sensitive to oxygen and moisture in the air. It is possible that the probe got exposed to oxygen/water and was oxidized before adding it to the alcohol-containing samples. This might explain the pink color seen. Also, this kit is very sensitive to methanol/ethanol/propanol vapor in the lab and the probe can react to this vapor and yield a pink color.
What is the recommended way to prepare and store the enzyme mix, i.e. do you vortex the enzyme mix or gently mix by inversion? Do you keep enzyme mix on ice or at room temperature? Should we use cold assay buffer to disolve enzyme mix?
On the day of the experiment, if all 96/100 samples+standards will be run together, the assay buffer warmed to RT can be used to reconstitute the enzyme mix. The mix can be vortexed and used for the assay immediately. If the enzymemix will be used multiple times, the reconstituted enzyme can be stored at 4C for up to 2 months. Alternately, the enzyme mix can be reconstituted in cold Assay buffer and kept on ice while the samples are being aliquoted into the wells. The reaction with the samples will occur at RT or 37C during the incubation in any case.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is essential to do the standards every time you do the expt to ensure there is no absorption of ethanol vapors into the reagents. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
What is the exact volume of sample required for this assay?
The kit detects 0.1-10 ppm alcohol (~10-800 nM) in samples. Hence depending upon the amount of ethanol in the samples, the volume needs to be optimized such that readings are within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Can protein content be used as an internal control for this assay?
Yes, a detergent-compatible BCA assay can be used for protein quantitation to normalize sample amount.
Sulie L. Chang, Binge-Like Exposure to Ethanol Enhances Morphine's Anti-nociception in B6 Mice. Front Psychiatry, Jan 2019; 30723430.
Wang, M. et al., (2017) Omega-3 polyunsaturated fatty acids ameliorate ethanol-induced adipose hyperlipolysis: A mechanism for hepatoprotective effect against alcoholic liver disease, Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, Aug.2017, https://doiorg/101016/jbbadis201708026
Zhang, Ning et al. (2017) Salvianolic acid B protects against chronic alcoholic liver injury via SIRT1-mediated inhibition of CRP and ChREBP in rats, Toxicol Lett. 2017 Feb 5;267:1-10.
Zhao et al., In vivo efficacy of HDL-like nanolipid particles containing multivalent peptide mimetics of apolipoprotein A-I. J. Lipid Res., Oct 2014; 55: 2053 - 2063.
Ye et al., Deletion of the Cardiolipin-specific Phospholipase Cld1 Rescues Growth and Life Span Defects in the Tafazzin Mutant: IMPLICATIONS FOR BARTH SYNDROME. J. Biol. Chem., Feb 2014; 289: 3114 - 3125.
For more citations of this product click here