Creating Optimal Conditions for OPA1 Isoforms by Western Blot in Skeletal Muscle Cells and Tissue.

Autor: Stephens DC; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.; Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA., Mungai M; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Crabtree A; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Beasley HK; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Garza-Lopez E; Department of Internal Medicine, University of Iowa, Iowa City, IA, 52242, USA., Neikirk K; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Bacevac S; Central Microscopy Research Facility, University of Iowa, Iowa City, IA, 52242, USA., Vang L; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Vue Z; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Vue N; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Marshall AG; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA., Turner K; Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA., Shao J; Central Microscopy Research Facility, University of Iowa, Iowa City, IA, 52242, USA., Murray S; Department of Cell Biology, College of Medicine, University of Pittsburgh, Pittsburgh, PA, 15260, USA., Gaddy JA; Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.; Tennessee Valley Healthcare Systems, U.S. Department of Veterans Affairs, Nashville, TN, 37212, USA., Wanjalla C; Vanderbilt University Medical Center: Department of Medicine, Division of Infectious Disease, Nashville, TN, USA., Davis J; Department of Biochemistry and Cancer Biology. Meharry Medical College, Nashville, TN, 37208, USA., Damo S; Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA., Hinton AO Jr; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.
Jazyk: angličtina
Zdroj: BioRxiv : the preprint server for biology [bioRxiv] 2023 Aug 19. Date of Electronic Publication: 2023 Aug 19.
DOI: 10.1101/2023.05.20.541601
Abstrakt: OPA1 is a dynamin-related GTPase that modulates various mitochondrial functions and is involved in mitochondrial morphology. There are eight different isoforms of OPA1 in humans and five different isoforms in mice that are expressed as short or long-form isoforms. These isoforms contribute to OPA1's ability to control mitochondrial functions. However, isolating OPA1 all long and short isoforms through western blot has been a difficult task. To address this issue, we outline an optimized western blot protocol to isolate 5 different isoforms of OPA1 on the basis of different antibodies. This protocol can be used to study changes in mitochondrial structure and function.
Databáze: MEDLINE