常见的固定-通透方法会导致一种II型跨膜蛋白出现人为定位。

Common fixation-permeabilization methods cause artifactual localization of a type II transmembrane protein.

作者信息

Benyair Ron, Lederkremer Gerardo Z

机构信息

Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.

Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel

出版信息

Microscopy (Oxf). 2016 Dec;65(6):517-521. doi: 10.1093/jmicro/dfw035. Epub 2016 Sep 1.

DOI:10.1093/jmicro/dfw035

PMID:27587511

Abstract

We found that a localization artifact can arise from common immunofluorescence methods. Specifically, cell fixation and permeabilization can cause mislocalization of a type II membrane-bound protein, ER mannosidase I, from its native localization in vesicles to the Golgi complex. Live cell microscopy and interestingly also mild cell fixation with paraformaldehyde without membrane permeabilization do not present this artifact.

摘要

我们发现，常见的免疫荧光方法可能会产生定位假象。具体而言，细胞固定和通透处理可导致一种II型膜结合蛋白——内质网甘露糖苷酶I——从其在囊泡中的天然定位错误定位于高尔基体复合体。活细胞显微镜观察，有趣的是，用多聚甲醛进行的轻度细胞固定且不进行膜通透处理时，不会出现这种假象。