Creating Optimal Western Blot Conditions for OPA1 Isoforms in Skeletal Muscle Cells and Tissue
Margaret Mungai, Amber Crabtree, Han Le, Johnathan Moore, Desiree Nguyen, Benjamin Rodriguez, Chanel Harris, Dominique C. Stephens, Heather K. Beasley, Edgar Garza-Lopez, Kit Neikirk, Bryanna Shao, Ashton Oliver, Genesis Wilson, Serif Bacevac, Larry Vang, Zer Vue, Neng Vue, Andrea G. Marshall, Kyrin Turner, Elma Zaganjor, Jianqiang Shao, Sandra Murray, Jennifer A. Gaddy, Celestine Wanjalla, Jamaine Davis, Steven M. Damo, Lori D. Banks, Antentor Hinton Jr
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Abstract
OPA1 is a dynamin-related GTPase that modulates mitochondrial dynamics and cristae integrity. Humans carry eight different isoforms of OPA1 and mice carry five, all of which are expressed as short- or long-form isoforms. These isoforms contribute to OPA1's ability to control mitochondrial energetics and DNA maintenance. However, western blot isolation of all long and short isoforms of OPA1 can be difficult. To address this issue, we developed an optimized western blot protocol based on improving running time to isolate five different isoforms of OPA1 in mouse cells and tissues. This protocol can be applied to study changes in mitochondrial structure and function. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Basic Protocol : Western Blot Protocol for Isolating OPA1 Isoforms in Mouse Primary Skeletal Muscle Cells
建立骨骼肌细胞和组织中OPA1亚型的最佳Western Blot条件。
OPA1是一种与动力蛋白相关的GTPase，可调节线粒体动力学和嵴完整性。人类携带8种不同的OPA1亚型，小鼠携带5种，它们都以短型或长型亚型的形式表达。这些同工异构体有助于OPA1控制线粒体能量和DNA维持的能力。然而，western blot分离所有的长和短亚型的OPA1可能是困难的。为了解决这一问题，我们开发了一种基于改进运行时间的优化western blot方案，以分离小鼠细胞和组织中的五种不同的OPA1亚型。该方案可用于研究线粒体结构和功能的变化。©2025作者。当前协议由Wiley期刊有限责任公司发表。基本协议：Western Blot分离小鼠原代骨骼肌细胞中OPA1亚型的协议。
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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