{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:47:28Z","timestamp":1782280048692,"version":"3.54.5"},"posted":{"date-parts":[[2023,5,22]]},"group-title":"Cell Biology","reference-count":17,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,8,19]]},"abstract":"<jats:title>Summary<\/jats:title>\n                <jats:p>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\u2019s 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.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Tweetable Abstract<\/jats:title>\n                  <jats:p>Western blot protocol optimization to visualize OPA1 isoforms.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Highlights<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>Protocol for isolating OPA1 isoforms in primary skeletal muscle myoblast and myotubes<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Steps for running isolated skeletal muscle cells from muscle tissue on a gel<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>How to collect samples in preparation for western blotting<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Detection of OPA1 isoforms<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" fig-type=\"figure\" orientation=\"portrait\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"541601v2_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                  <jats:table-wrap id=\"utbl1\" orientation=\"portrait\" position=\"float\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"541601v2_utbl1\" position=\"float\" orientation=\"portrait\"\/>\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"541601v2_utbl1a\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:table-wrap>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Method Summary<\/jats:title>\n                  <jats:p>Samples for western blot analysis are isolated from lysed cells, loaded onto a gel, and ran using optimized conditions to better isolate OPA1 isoforms. Samples are transferred to a membrane for incubation and protein detection using OPA1 antibodies.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.05.20.541601","type":"posted-content","created":{"date-parts":[[2023,5,22]],"date-time":"2023-05-22T10:25:11Z","timestamp":1684751111000},"source":"Crossref","is-referenced-by-count":1,"title":["Creating Optimal Conditions for OPA1 Isoforms by Western Blot in Skeletal Muscle Cells and Tissue"],"prefix":"10.64898","author":[{"given":"Dominique C.","family":"Stephens","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margaret","family":"Mungai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amber","family":"Crabtree","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather K.","family":"Beasley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edgar","family":"Garza-Lopez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kit","family":"Neikirk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serif","family":"Bacevac","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larry","family":"Vang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zer","family":"Vue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neng","family":"Vue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea G.","family":"Marshall","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyrin","family":"Turner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianqiang","family":"Shao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Murray","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer A.","family":"Gaddy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celestine","family":"Wanjalla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamaine","family":"Davis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven","family":"Damo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7730-952X","authenticated-orcid":false,"suffix":"Jr","given":"Antentor O.","family":"Hinton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2023082208301060000_2023.05.20.541601v2.1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvp181"},{"key":"2023082208301060000_2023.05.20.541601v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.06.025"},{"key":"2023082208301060000_2023.05.20.541601v2.3","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.bbamcr.2012.08.004","article-title":"The dynamin GTPase OPA1: More than mitochondria?","volume":"1833","year":"2013","journal-title":"Biochimica et Biophysica Acta (BBA) - Molecular Cell Research"},{"key":"2023082208301060000_2023.05.20.541601v2.4","doi-asserted-by":"publisher","DOI":"10.1038\/79936"},{"key":"2023082208301060000_2023.05.20.541601v2.5","doi-asserted-by":"publisher","DOI":"10.1038\/79944"},{"key":"2023082208301060000_2023.05.20.541601v2.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.05.073"},{"key":"2023082208301060000_2023.05.20.541601v2.7","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2008.05401.x"},{"key":"2023082208301060000_2023.05.20.541601v2.8","doi-asserted-by":"crossref","unstructured":"Gilkerson, R. , De La Torre, P. & St. Vallier, S. 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