一种用于评估巨噬细胞合成[³H]亮氨酸标记蛋白质的微系统。

A microsystem to evaluate the synthesis of [3H]leucine labeled proteins by macrophages.

作者信息

Varesio L, Eva A

出版信息

J Immunol Methods. 1980;33(3):231-8. doi: 10.1016/0022-1759(80)90210-0.

Abstract

A method is described for evaluating protein synthesis by adherent M phi by measuring the incorporation of [3H]leucine into TCA precipitable material. By using guanidine-HCl it was possible to remove completely the radiolabeled proteins from the adherent cells that were cultured in microwells, and retain TCA precipitable material. This procedure enabled us to harvest the TCA precipitable proteins with a semiautomatic cell harvester. The guanidine-HCl treatment did not affect the recovery of the radioactive proteins and did not alter the sensitivity of the assay. This method is very simple and rapid and, since it is suitable for processing microcultures, permits detailed studies on the biology of small numbers of M phi.

摘要

描述了一种通过测量[3H]亮氨酸掺入三氯乙酸（TCA）可沉淀物质来评估贴壁巨噬细胞（M phi）蛋白质合成的方法。通过使用盐酸胍，能够从微孔中培养的贴壁细胞中完全去除放射性标记的蛋白质，并保留TCA可沉淀物质。该程序使我们能够使用半自动细胞收集器收获TCA可沉淀蛋白质。盐酸胍处理不影响放射性蛋白质的回收率，也不改变测定的灵敏度。该方法非常简单快速，并且由于适用于处理微量培养物，因此可以对少量M phi的生物学进行详细研究。