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Alcohol Dehydrogenase - Assay
Source:
Yeast
CAS:
9031-72-5
EC:
1.1.1.1
Alcohol Dehydrogenase Assay
Method
The reaction velocity is determined by the method of A Vallee and Hoch (1955) in which the rate of absorbance at 340 nm resulting from reduction of NAD is measured. One unit reduces one micromole of NAD per minute at 25°C under the specified conditions.
Reagents
- 0.1 M Sodium pyrophosphate buffer, pH 9.2
- 2 M Ethanol. Dilute 12.12 ml of 95% ethanol to 100 ml with reagent grade water.
- 0.025 M NAD. Note: NAD may vary in salt form and degree of hydration. Care should be exercised to use an analytical grade and the correct molecular weight.
- 0.1 M Phosphate buffer, pH 7.5
- 0.1% Albumin (BSA)
Enzyme
Dissolve lyophilized enzyme at one mg/ml in 0.1 M phosphate buffer, pH 7.5.
Immediately prior to use, dilute all enzymes to a concentration of 0.05-0.25 units/ml in 0.1% albumin.
For protein determinations, dissolve enzyme in 0.1 M phosphate buffer, pH 7.5.
Procedure
Set spectrophotometer to 340 nm and 25°C.
Pipette into each cuvette:
0.1 M Pyrophosphate buffer 1.5 ml 2.0 M Ethanol 0.5 ml 0.025 M NAD 1.0 ml
Incubate in spectrophotometer for 3-4 minutes at 25°C to achieve temperature equilibrium and establish blank rate, if any. At zero time, add 0.1 ml of appropriately diluted enzyme to the cuvette and record the A340 for 3-4 minutes. Calculate the ΔA340/minute from the initial linear portion of the curve.
Calculation
Alcohol Dehydrogenase Products
Description
Activity
Code
Cat. #
Size
Price
Description
Alcohol Dehydrogenase, Suspension
Source:
Yeast
Two times crystallized. A suspension in 2.4M ammonium sulfate containing 3% pyrophosphate and 0.1% glycine.
Store at 2-8°C.
Ice Pack required
Code
ADHS
Description
Alcohol Dehydrogenase, Lyophilized
Source:
Yeast
Two times crystallized. A lyophilized powder.
Store at -20°C.
Ice Pack required
Code
ADHL
Product details
LS001069
100 mg
$115.00
LS001070
1 gm
$860.00
LS001071
Bulk
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