Optimizing In Situ Proximity Ligation Assays for Mitochondria, ER, or MERC Markers in Skeletal Muscle Tissue and Cells

优化骨骼肌组织和细胞中线粒体、ER 或 MERC 标记物的原位邻近连接检测

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作者:Amber Crabtree, Han Le, Chanel Harris, Ashton Oliver, Andy Barillas, Johnathan Moore, Desiree Ngoc-Ha Nguyen, Izabella Marie Rabago, Benjamin Rodriguez, Dominique C Stephens, Heather K Beasley, Edgar Garza-Lopez, Kit Neikirk, Margaret Mungai, Larry Vang, Zer Vue, Neng Vue, Andrea G Marshall, Kyrin T

Abstract

Proximity ligation assays (PLAs) use specific antibodies to detect endogenous protein-protein interactions. PLAs are a highly useful biochemical technique that allow two proteins within proximity to be visualized with fluorescent probes amplified by PCR. While this technique has gained prominence, the use of a PLA in mouse skeletal muscle (SkM) is novel. In this article, we discuss how the PLA method can be used in SkM to study the protein-protein interactions within mitochondria-endoplasmic reticulum contact sites (MERCs). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Proximity ligation assay for skeletal muscle tissue and myoblast for MERC proteins.

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