通过跟踪pH指示剂的吸光度变化来测定酶活性。

Assay for enzyme activity by following the absorbance change of pH-indicators.

作者信息

Yao Y, Wang M H, Zhao K Y, Wang C C

机构信息

National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing, China.

出版信息

J Biochem Biophys Methods. 1998 Jun 11;36(2-3):119-30. doi: 10.1016/s0165-022x(98)00005-0.

DOI:10.1016/s0165-022x(98)00005-0

PMID:9711498

Abstract

Based on the absorbance change of indicators with the concentration of hydrogen ion released from an enzyme-catalyzed reaction, a convenient colorimetric method was established for the assay of acidic phospholipase A2 and glycogen phosphorylase b. Brilliant yellow and bromothymol blue were chosen as indicators for assays of acidic phospholipase A2 and glycogen phosphorylase b by following the absorbance changes at 495 and 615 nm, respectively. The method is simple, sample-saving, sensitive and valid for a wide range of enzyme concentrations. It can be extended for assaying other enzymes catalyzing reactions with hydrogen ion concentration changes.

摘要

基于指示剂吸光度随酶催化反应释放的氢离子浓度的变化，建立了一种简便的比色法用于测定酸性磷脂酶A2和糖原磷酸化酶b。分别选择亮黄和溴百里酚蓝作为指示剂，通过跟踪495 nm和615 nm处的吸光度变化来测定酸性磷脂酶A2和糖原磷酸化酶b。该方法简单、节省样品、灵敏且对广泛的酶浓度有效。它可扩展用于测定其他催化伴随氢离子浓度变化反应的酶。