Michael D Roberts, Kaelin C Young, Carlton D Fox, Christopher G Vann, Paul A Roberson, Shelby C Osburn, Johnathon H Moore, Petey W Mumford, Matthew A Romero, Darren T Beck, Cody T Haun, Veera L D Badisa, Benjamin M Mwashote, Victor Ibeanusi, Andreas N Kavazis
{"title":"分离啮齿动物和人类骨骼肌肌浆蛋白和肌原纤维蛋白的优化程序。","authors":"Michael D Roberts, Kaelin C Young, Carlton D Fox, Christopher G Vann, Paul A Roberson, Shelby C Osburn, Johnathon H Moore, Petey W Mumford, Matthew A Romero, Darren T Beck, Cody T Haun, Veera L D Badisa, Benjamin M Mwashote, Victor Ibeanusi, Andreas N Kavazis","doi":"10.14440/jbm.2020.307","DOIUrl":null,"url":null,"abstract":"<p><p>Several published protocols exist for isolating contractile or myofibrillar (MF) proteins from skeletal muscle, however, achieving complete resuspension of the myofibril pellet can be technically challenging. We performed several previously published MF isolation methods with the intent of determining which method was most suitable for MF protein isolation and solubilization. Here, we provide an optimized protocol to isolate sarcoplasmic and solubilized MF protein fractions from mammalian skeletal muscle suitable for several downstream assays.</p>","PeriodicalId":73618,"journal":{"name":"Journal of biological methods","volume":"7 1","pages":"e127"},"PeriodicalIF":0.0000,"publicationDate":"2020-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/5a/c7/jbm-7-1-e127.PMC7081056.pdf","citationCount":"16","resultStr":"{\"title\":\"An optimized procedure for isolation of rodent and human skeletal muscle sarcoplasmic and myofibrillar proteins.\",\"authors\":\"Michael D Roberts, Kaelin C Young, Carlton D Fox, Christopher G Vann, Paul A Roberson, Shelby C Osburn, Johnathon H Moore, Petey W Mumford, Matthew A Romero, Darren T Beck, Cody T Haun, Veera L D Badisa, Benjamin M Mwashote, Victor Ibeanusi, Andreas N Kavazis\",\"doi\":\"10.14440/jbm.2020.307\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Several published protocols exist for isolating contractile or myofibrillar (MF) proteins from skeletal muscle, however, achieving complete resuspension of the myofibril pellet can be technically challenging. We performed several previously published MF isolation methods with the intent of determining which method was most suitable for MF protein isolation and solubilization. Here, we provide an optimized protocol to isolate sarcoplasmic and solubilized MF protein fractions from mammalian skeletal muscle suitable for several downstream assays.</p>\",\"PeriodicalId\":73618,\"journal\":{\"name\":\"Journal of biological methods\",\"volume\":\"7 1\",\"pages\":\"e127\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-02-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/5a/c7/jbm-7-1-e127.PMC7081056.pdf\",\"citationCount\":\"16\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of biological methods\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.14440/jbm.2020.307\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2020/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of biological methods","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.14440/jbm.2020.307","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2020/1/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
An optimized procedure for isolation of rodent and human skeletal muscle sarcoplasmic and myofibrillar proteins.
Several published protocols exist for isolating contractile or myofibrillar (MF) proteins from skeletal muscle, however, achieving complete resuspension of the myofibril pellet can be technically challenging. We performed several previously published MF isolation methods with the intent of determining which method was most suitable for MF protein isolation and solubilization. Here, we provide an optimized protocol to isolate sarcoplasmic and solubilized MF protein fractions from mammalian skeletal muscle suitable for several downstream assays.