About Calzyme Contact Us Locations Our Policies

  Welcome to Calzyme!

Alcohol Dehydrogenase (ADH)

(Alcohol:NAD+ oxidoreductase; EC

Alcohol dehydrogenase (ADH) catalyzes the oxidation of alcohol and the reduction of aldehydes as shown below:

The enzyme occurs in various mammalian tissues but is found in relatively high concentrations in liver and kidney. It is a zinc-containing enzyme and is activated by glutathione and EDTA and inhibited by heavy metals. ADH from equine liver has a molecular weight of 80,000, while the yeast enzyme has a molecular weight of 141,000.

Alcohol dehydrogenase is widely used for the determination of ethanol in biological fluids. It can also be used in coupled enzyme reactions for determination of metabolites in biological fluids. It's optimum pH is 8.6 to 9.0 and it has an extension coefficient of 12.6 and isoelectric point is 5.4. The enzyme is activated with sulfhydryl reagents, like mercapethanol and dithiotreitol.

1.Source: Bakers Yeast
   Form: Freeze-dried powder 
   Solubility: Distilled water or dilute buffer 
   Stability: Store at -20 C (-4 F) 
   Activity (approx.): 300 U/mg protein 
   Protein (approx.): 90-95% (biuret) 
   Catalog No.: 054A0300 

The amount of enzyme which catalyzes the reduction of one micromole NAD+ per minute at 25C and pH 8.8.

The increase in absorbance at 340 nm caused by the reduction of NAD+, is a measure of the catalytic activity of ADH (Vallee, B.L. and Hoch, F.L., Proc. Nat. Acad. Sci. USA, 41, 327, 1955).

  1. 0.05 M Sodium pyrophosphate buffer, pH 8.8.
  2. 96% Ethanol (substrate).
  3. 0.025 M NAD+ (16.7 mg/ml) in 0.01 M Tris-HCL buffer, pH 7.5. Prepare fresh.
  4. 0.01 M Tris-HCL buffer, pH 7.5, containing 0.1% bovine serum albumin (BSA).
  5. Alcohol dehydrogenase (enzyme) - Dissolve sufficient amount of enzyme in 0.01 M Tris-HCL buffer containing 0.1% BSA, pH 7.5, to give a concentration of 0.1-0.5 U/ml. Prepare fresh immediately prior to assay.

  1. Set spectrophotometer (equipped with temperature control) at 340 nm and 25C.
  2. In a cuvette pipette the following reagents in the amounts indicated:
    Sodium pyrophosphate buffer      1.4 ml
    NAD+                             1.4 ml
    Ethanol (substrate)              0.1 ml
    Incubate cuvette in spectrophotometer, at 25C for 5 min. to achieve temperature equilibration and then record absorbance at 340 nm (blank).
  3. Initiate the reaction by adding 0.1 ml of ADH (enzyme) solution to the cuvette. Record the increase in absorbance at 340 nm for 5 min.
  4. Calculate the (delta)E340nm/min

Activity (U/mg) = (ΔE340nm/min)(Total Vol.)(Enz. Diln.)
(6.22)(Enz. Vol.)(mg Enz./ml)

©1999-2018 Calzyme All Rights Reserved powered by corezon
All logos and trademarks are properties of their respective companies. Product price, specification, warranty and availability subject to change without notice.
2-Phosphoenol Pyruvate Acid 3-Indoxyl Phosphate
3-Phosphoglycerate Kinase Acetone Powders
Acetyl Cholinesterase Acid Phosphatase
Actin Acylase
Adenosine 5'-diphosphate Adenosine 5'-monophosphate (AMP)
Adenosine 5'-triphosphate (ATP) Adenosine Deaminase
Alcohol Dehydrogenase Aldehyde Dehydrogenase
Aldolase Alkaline Phosphatase
alpha-Amylase alpha-D-Glucose-1-Phosphate
alpha-Glucosidase alpha-Ketoglutarate
Amylopectin Amylose
Apo-d-Amino Acid Oxidase Ascorbate Oxidase
Avidin beta-Amylase
beta-Glucosidase beta-Glucronidase
beta-NAD beta-NADH
beta-NADPH Bromelain
Butyryl Cholinesterase C-Reactive Protein
Carbonic Anhydrase Carboxypeptidase A
Carboxypeptidase B Carboxypeptidase Y
Catalase Catechol-o-Methyl Transferase
Cathepsin D Cholesterol Esterase
Chymotrypsin d-Amino Acid Oxidase
d-Glucose-6-Phosphate Deoxyribonuclease (DNase)
Deoxyribonucleic Acid (DNA) Elastase
Enterokinase Ferritin
G-6-phosphate Dehydrogenase gamma-Glutamyl Transferase
Glucose Oxidase Glutamate Dehydrogenase
Glutathione Peroxidase Glycerol Kinase
Glycerol-3-phosphate Dehydrogenase Glycosylated Albumin
Glycosylated Hemoglobin GOT (AST)
GPT (ALT) Hemoglobin
 Hexokinase Histamine-n-Methyl Transferase
Hyaluronic Acid Hyaluronidase
Insulin L-Arginase
L-gamma-Glutamide Nitroanilide Lactate Dehydrogenase
Lactoperoxidase Leucine Aminopeptidase
Lipase Lipoamide Dehydrogenase
Lipoxidase Luciferase
Lysozyme Malate Dehydrogenase
Mutarotase Myelin
Myoglobin Myosin
Oxyhemoglobin p-Nitrophenyl Phosphate
Pepsin Peroxidase
Phosphorylase-b Phspho(enol) Pyruvate Carboxylase
PNMT Prophospholipase A2
Protein Methylase II Pyruvate Kinase
Renin Rennin
Ribonuclease Sedoheptulose
Stachyose Superoxide Dismutase
Thyroglobulin Triose Phosphate Isomerase
Trypsin Trypsin Inhibitor
Tyrosinase Urease
Uricase Xanthine Oxidase