A simple and highly reproducible method for the detection of erythrocyte membrane ZIP metal transporters by immunoblotting

一种简单且高度可重复的免疫印迹检测红细胞膜 ZIP 金属转运体的方法

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作者:Varalakshmi Vungutur, Suetmui Yu, Shannon McCabe, Caitlin Fung, Ningning Zhao

Abstract

Manganese is one of the essential trace elements found in erythrocytes. Metal transporters situated on the plasma membrane generally facilitate the movement of manganese into and out of cells. This study aims at determining whether two recently discovered manganese importers, ZIP8 and ZIP14, are located in the erythrocyte membrane. We outline a simple, effective and repeatable method for the isolation of erythrocyte membrane from a minimum of 50 µL mouse blood, followed by the identification of ZIP metal transporters using immunoblotting. Our results revealed that ZIP8 is expressed within the erythrocyte membrane, in contrast to ZIP14 which is not identified using immunoblotting approach. A direct measurement of the ZIP8 protein expression in erythrocyte membranes could provide valuable information for further analyzing its biological function.

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