/
/
Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
É necessária uma assinatura da JoVE para visualizar este conteúdo. Faça login ou comece sua avaliação gratuita.
Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
8,329 Views
•
08:31 min
•
October 03, 2018
DOI:
Summary
Automatically generated
Here we present a protocol for anaerobic protein purification, anaerobic protein concentration, and subsequent kinetic characterization using an oxygen electrode system. The method is illustrated using the enzyme DesB, a dioxygenase enzyme which is more stable and active when purified and stored in an anaerobic environment.
Explore More Videos
AnaerobicProtein PurificationKinetic AnalysisOxygen ElectrodeDesB DioxygenaseEnzyme ActivityInhibitionInorganic BiochemistryOxygen-sensitive MetalsProtein ExpressionCell PelletAmylose ColumnLysozymeHigh-pressure HomogenizationNitrogen GasGlove BoxAmylose Column BufferElution BufferFerrous Ammonium SulfateDithiothreitol
Read Article
Cite this Article
Uchendu, S. N., Rafalowski, A., Cohn, E. F., Davoren, L. W., Taylor, E. A. Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition. J. Vis. Exp. (140), e58307, doi:10.3791/58307 (2018).
Copy