Maita E, Endo Y, Ogura Y
Anal Biochem. 1983 Jan;128(1):36-40. doi: 10.1016/0003-2697(83)90340-8.
A simple method for the assay of bradykinin (BK)-degrading enzymes was investigated. The procedure of the method includes enzymatic degradation of BK, separation of the residual BK on a small P-cellulose column (0.6 X 3 cm), and its fluorometrical determination based on the reaction with fluorescamine. BK was separated completely from its fragments produced during enzymatic reaction by the column chromatography. The recovery rate of BK was 96 +/- 3%. Quantitative determinations could be carried out on 0.2 nmol of BK, at least in the fluorometry. This method was available for the assay of the enzymes in tissue homogenates as well as in purified preparations, and its usefulness for the study of the enzymes is presented.
研究了一种测定缓激肽(BK)降解酶的简单方法。该方法的步骤包括BK的酶促降解、在小型P-纤维素柱(0.6×3 cm)上分离残留的BK以及基于与荧光胺反应的荧光测定法。通过柱色谱法可将BK与其在酶促反应过程中产生的片段完全分离。BK的回收率为96±3%。至少在荧光测定法中,对0.2 nmol的BK即可进行定量测定。该方法可用于测定组织匀浆以及纯化制剂中的酶,并展示了其在酶研究中的实用性。