Acidimetric assay for phospholipase A using egg yolk suspension as substrate.
A convenient acidimetric assay for phospholipase A using egg yolk suspension as substrate has been developed. The substrate mixture consists of 1 part egg yolk, 1 part 8.1 mM sodium deoxycholate, and 1 part 18 mM calcium chloride. Phospholipase A activity is measured by following the initial rate of pH change, which is linear between pH 8.0 and 7.75 and is proportional to enzyme concentration over a wide range. The assay is highly reproducible, with a coefficient of variation of 3%, and as sensitive as most established assays for phospholipase A. The assay uses inexpensive and easily available substrate and is simple to perform. It is particularly useful for monitoring phospholipase A activity in chromatography fractions.
Tan
N H
NH
Department of Biochemistry, University of Malaya, Kuala Lumpur.
Tan
C S
CS
D000818:Animals; D002645:Chickens; D002845:Chromatography; D004530:Egg Yolk; D004546:Elapid Venoms / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D010179:Pancreas / Q000201:enzymology; D010740:Phospholipases / Q000032:analysis*; D010741:Phospholipases A / Q000032:analysis*; D013379:Substrate Specificity; D013552:Swine
D004546:Elapid Venoms; D010740:Phospholipases; D010741:Phospholipases A