使用125I标记酚酞对微粒体UDP-葡萄糖醛酸基转移酶进行放射性测定。

The radiometric assay of microsomal UDP-glucuronyltransferase using 125I-labelled phenolphthalein.

作者信息

Aw S E

出版信息

Clin Chim Acta. 1975 Aug 18;63(1):73-80. doi: 10.1016/0009-8981(75)90381-2.

Abstract

A rapid and sensitive method is described for assaying UDP-glucuronyl-transferase (EC 2.4.1.17) activity. The substrate is the relatively apolar phenolphthalein, labelled with 125I on one of its two phenolic rings leaving a free hydroxyl group for glucuronidation. Extraction with ethyl acetate leaves the glucuronide in the aqueous reaction medium which is then counted. Less than 10 mug microsomal protein is required and the glucuronidation of 10 ng 125I-labelled phenolphthalein (0.03 nmoles) is easily detected.

摘要

本文描述了一种快速灵敏的方法，用于测定UDP-葡萄糖醛酸基转移酶（EC 2.4.1.17）的活性。底物是相对非极性的酚酞，在其两个酚环之一上用125I标记，留下一个游离羟基用于葡萄糖醛酸化。用乙酸乙酯萃取后，葡萄糖醛酸化物留在水性反应介质中，然后进行计数。所需微粒体蛋白少于10微克，10纳克125I标记的酚酞（0.03纳摩尔）的葡萄糖醛酸化很容易检测到。