• Product name

    Ethanol Assay Kit
  • Detection method

  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay type

  • Sensitivity

    > 10 nM
  • Range

    2 µM - 200 µM
  • Assay time

    1h 00m
  • Product overview

    Ethanol Assay Kit ab65343 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.

    In the ethanol assay protocl, alcohol oxidase oxidizes ethanol to generate H2O2 which reacts with a probe to generate color (absorbance max 570 nm) and fluorescence (Ex/Em 535/587 nm).

    The kit detects 0.1-10 ppm alcohol (~10-800 nM).

    Ethanol assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 30 min at 37ºC
    - analyze with microplate reader

  • Notes

    PLEASE NOTE: Extreme care should be taken to ensure that no alcohol vapors (ethanol, methanol, propanol) are in the laboratory air where this assay is to be performed. Alcohol vapors in the air will be rapidly absorbed by kit components resulting in very high background making the kit unusable. 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.

  • Platform

    Microplate reader


  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Ethanol Assay Buffer WM 1 x 25ml
    Ethanol Enzyme Mix Green 1 vial
    Ethanol Probe (in DMSO, anhydrous) Red 1 x 200µl
    Ethanol Standard Yellow 1 x 500µl
  • Research areas

  • Relevance

    Alcohol (ethanol C2H5OH) is a clear colorless liquid rapidly absorbed from the gastrointestinal tract and distributed through the body. Ethanol has a depressive effect on the central nervous system and because of its psychoactive effects, it is considered a drug. Ethanol is metabolized by the body as an enerfy-providing carbohydrate nutrient, as it metabolized into acetyl CoA, an intermediate common with glucose than can be used for energy in the citric acid cycle (TCA) or for biosynthesis. Ethanol within the human body is converted into acetaldehyde by alcohol dehydrogenase and then into acetic acid by acetaldehyde dehydriogenase. Acetaldehyde, the product of the first step of the alcohol breakdown, is linked to most of the toxic clinical effects of alcohol.
  • Alternative names

    • Ethyl alcohol


  • Ethanol in blood plasma was analyzed using ethanol assay kit (ab65343). Ethanol concentrations were determined 60 minutes after urethane administration in P0/1 mice and rats. 

  • 0.5 nmol of ethanol was added to diluted human biological fluids (1/10) and spiked samples were measured fluorimetrically (background subtracted, duplicates; +/- SD).

  • 5 nmol of ethanol was added to diluted human biological fluids (1/10) and spiked samples were measured colourimetrically (background subtracted, duplicates; +/- SD).

  • Standard curves with background signal subtracted (duplicates; +/- SD).

  • Ethanol Standard Curve. Ethanol standard was diluted as described and the assay was performed according to the kit instructions.



This product has been referenced in:

  • Cananzi SG & Mayhan WG In Utero Exposure to Alcohol Impairs Reactivity of Cerebral Arterioles and Increases Susceptibility of the Brain to Damage Following Ischemia/Reperfusion in Adulthood. Alcohol Clin Exp Res 43:607-616 (2019). Read more (PubMed: 30748017) »
  • McCarter KD  et al. Influence of low-dose alcohol consumption on post-ischemic inflammation: Role of cystathionine ?-lyase. Alcohol 76:81-89 (2019). Read more (PubMed: 30597416) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A


The expiry date of the kits is 1 year from date of purchase. Please discuss the expiry date with our distributor in Russia. http://www.almabion.ru/

Please note that the guarantee is only valid when the components of the kit are stored at correct temperature as suggested in the kit protocol booklet. Please check the booklet.

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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.

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Gracias por contactarnos.

Siempre y cuando el suero se alicuotée y congele, evitando ciclos de congelación/descongelación, no debe haber ningún problema. No hay una cantidad determinada de suero requerido para hacer el ensayo. Lo que es importante es hacer varias diluciones del suero para que el rango de alcohol a testar caiga dentro del rango lineal de la curva estándar.

Los valores de alcohol en suero suelen variar entre ˜ 0.01-0.16% w/v, por lo que se sugieren diluciones dentro del rango 1:10 – 1:100, utilizando después 10-30ul de estas diluciones.

Como bien mencionas, es importante además proteger las muestras de la luz y de los vapores del laboratorio que pueden distorsionar las medidas.

Espero que esta información sea de utilidad. Si tienes cualquier otra consulta no dudes en volvernos a contactar.

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Yes, you can use this kit to assay mouse samples. The assay will measure ethyl alcohol in cell culture supernatant, urine, serum, plasma, other biological fluids, and tissue extracts regardless of the species of the samples.

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Suite à notre conversation téléphonique, j’ai eu une réponse de laboratoire qui pense que ce kit pourrait donner de bons résultants chez la souris bien qu’il ne l’ait pas testé auparavant. Je vous suggère de regarder les références ci-dessous pour voir comment d’autres chercheurs ont utilisé des kits similaires.

Ethanol Assay Kit

- Gardner, J. et. al. Requirement of the Type II Secretion System for Utilization of Cellulosic Substrates by Cellvibrio japonicas.Appl. Envir. Microbiol., 2010; 76: 5079-5087.

- Hsu D-Z et al (2009) JPEN J Parenter Enteral Nutr; 33: 423 - 427.

- Li, H. et. al. The Small GTPase RacA Mediates Intracellular Reactive Oxygen Species Production, Polarized Growth, and Virulence in the Human Fungal Pathogen Aspergillus fumigatus. Eukaryot. http://ec.asm.org/cgi/content/abstract/10/2/174?maxtoshow=&HITS=10&hits=100&RESULTFORMAT=1&andorexacttitle=and&andorexacttitleabs=and&fulltext=Biovision&andorexactfulltext=and&searchid=1&FIRSTINDEX=0&sortspec=match&fdate=2/1/2011&tdate=2/28/2011&resourcetype=HWCIT

- Li, H. et. al. The Small GTPase RacA mediates intracellular reactive oxygen species production, polarized growth, and virulence in the human fungal pathogen Aspergillus fumigatus. http://ec.asm.org/cgi/content/abstract/EC.00288-10v1?maxtoshow=&HITS=10&hits=60&RESULTFORMAT=1&andorexacttitle=and&andorexacttitleabs=and&fulltext=BioVision&andorexactfulltext=and&searchid=1&FIRSTINDEX=0&sortspec=match&fdate=12/1/2010&resourcetype=HWCIT

- Sur, K. et al. Immiscible Phase Nucleic Acid Purification Eliminates PCR Inhibitors with a Single Pass of Paramagnetic Particles through a Hydrophobic Liquid. J. Mol. Diagn., Sep 2010; 12: 620 - 628.

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Thank you for your recent telephone enquiry.

I can confirm that serum from such samples made using heparin would be compatible with ab65343 Ethanol Assay Kit.

You can store this serum in aliquots for a couple of weeks at -80oC before analysis.

I hope this information has been helpful for you. If you have any further questions, please do not hesitate to contact us.

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Unfortunately we do not have a kit to assay for methanol conc.

Although I have not come across any instances in which the ethanol assay kit is used for methanol, it may be theoretically possible. The ethanol assay kit principle involves the conversion of ethanol to H2O2 by alcohol oxidase. This oxidase can also act on methanol to generate H2O2. Therefore it looks likely that the customer can use the ethanol assay kit for methanol, but some slight optimization/standardization might be in order for most efficient results.

Hope this information has been helpful for you. Please let me know if you have any other questions.

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Thank you for your patience. The lab has informed me that the extinction coefficient for the ethanol probe has not been determined. I have attached the absorption and emissions spectra for the probe. If you could let me know why you need this information, I will be happy to see if we can find an alternative way to help answer your question?

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Thank you for contacting us yesterday in regards the Ethanol Assay Kit (ab65343).

As discussed over the phone, I can now confirm that using the excitation wavelength of 544 nm instead of the recommended 535 should be fine as long as the machine’s bandwidth of range of detection has a flexibility of ±10. If this is not the case, measurement at 544 nm when the recommended wavelength is 535 nm will result in slightly lower and possibleinefficient signals.The relative levels will most likely remain the same, but the absolute detection values will be compromised.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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Thank you for your recent telephone enquiry. I am sorry it has taken some time to confirm this information for you with our source on this occasion.

From this equation,I can confirm that Sa is the sample amount from the Standard Curve (nmol). You can do it in two ways.

1. You can either plot the OD value you get form the sample, plot it onto the standard curve on the Y-axis and extrapolate it to find the corresponding concentration on the x-axis to find the Sa.

2. You can use the equation y = mx+c. So substitute the OD value for y, the equation will give you the values for m and c and then you can find the value for x, which is the Sa.

You can then use the equation provided in the protocolto work out the concentration in your sample:

Concentration = Sa / Sv (nmol/ìl or mM)

I hope this information has been helpful for you. If you have any further questions, please do not hesitate to contact us.

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1-10 of 16 Abreviews or Q&A

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