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Phosphatase, Acid - Assay
Source:
Wheat Germ (Triticum vulgare)
CAS:
9001-77-8
EC:
3.1.3.2
Phosphatase, Acid Assay
Method
Based on the work of Brandenberger and Hanson (1953) and Hofstee (1954). The initial rate of hydrolysis of o-carboxyphenyl phosphate is determined by following the increase in absorbance at 300 nm resulting from the release of salicylic acid. One unit hydrolyzes one micromole of o-carboxyphenyl phosphate per minute at 25°C, pH 5.0 under the specified conditions. (An equivalent specific activity is obtained under above conditions using p-nitrophenyl-phosphate substrate).
Reagents
- 0.15 M Sodium acetate buffer, pH 5.0
- 3.65 mM o-Carboxyphenyl-phosphate
Enzyme
Dissolve at a concentration of 1.0 mg/ml in reagent grade water.
Procedure
Adjust the spectrophotometer to 300 nm and 25°C.
Pipette into each cuvette as follows:
0.15 M Acetate buffer, pH 5.0 2.0 ml 3.65 mM o-Carboxyphenyl-phosphate 0.5 ml
Incubate in spectrophotometer for 3-4 minutes to reach temperature equilibration and establish blank rate, if any. Add 0.5 ml enzyme and record increase in A300 for 4-5 minutes. Calculate ΔA300/minute from the initial linear portion of the curve.
Calculation
Phosphatase, Acid Products
Description
Activity
Code
Cat. #
Size
Price
Description
Phosphatase, Acid
Source:
Wheat Germ (Triticum vulgare)
A non-specific esterase partially purified to the 0.35-0.55 fraction by the method described by Singer, JBC, 174, 11 (1948). Also active as a lipase. A lyophilized powder.
Store at -20°C.
Code
AP
Product details
LS001141
1 gm
$130.00
LS001144
Bulk
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