A Simplified Approach for the Direct Detection of Protein Oxidation in Muscle Using In-Gel and Microplate Assays

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dc.contributor.advisorPerry, Christopher G. R.
dc.creatorNejatbakhsh, Ali
dc.date.accessioned2016-09-20T18:58:42Z
dc.date.available2016-09-20T18:58:42Z
dc.date.copyright2016-05-19
dc.date.issued2016-09-20
dc.date.updated2016-09-20T18:58:42Z
dc.degree.disciplineKinesiology & Health Science
dc.degree.levelMaster's
dc.degree.nameMSc - Master of Science
dc.description.abstractReactive oxygen species (ROS) are emerging as regulators of protein redox states which influence many physiological processes. Current methods for the detection of protein redox states, including mass spectrometry, are expensive and not easily accessible or poorly validated. IRdye800CW Maleimide, a highly sensitive maleimide-based infrared dye, has been reported to detect the redox state of immunoprecipitated proteins in cardiac muscle using modified western blot procedures, but has yet to be validated as a novel tool to detect redox conditions throughout the proteome using common and cost-effective assays. In this study we tested the efficacy of IRDye800CW Maleimide in detecting protein redox state in cardiac muscle using mini-gel SDS-PAGE and an in-well approach. Chemical and heat-induced modifications to cardiac tissue redox states were accurately detected with the dye in cardiac lysates. These findings were also validated against HPLC measures of GSSG content.
dc.identifier.urihttp://hdl.handle.net/10315/32347
dc.language.isoen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectKinesiology
dc.subject.keywordsOxidation
dc.subject.keywordsProtein
dc.subject.keywordsIn-gel
dc.subject.keywordsMicroplate
dc.subject.keywordsSignaling
dc.subject.keywordsRedox
dc.subject.keywordsRedox state
dc.subject.keywordsIrdye800cw maleimide
dc.subject.keywordsMethods
dc.subject.keywordsRedox biology
dc.subject.keywordsOxidation
dc.subject.keywordsReduction
dc.subject.keywordstcep
dc.subject.keywordsh2o2
dc.subject.keywordsgsh
dc.subject.keywordsgssg
dc.subject.keywordsgsh:gssg
dc.subject.keywordsCarbonylation
dc.subject.keywordsOxidative stress
dc.subject.keywordsCysteine
dc.subject.keywordsThiol
dc.titleA Simplified Approach for the Direct Detection of Protein Oxidation in Muscle Using In-Gel and Microplate Assays
dc.typeElectronic Thesis or Dissertation

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