Anaerobic Protein Concentration, Buffer Exchange, and DesB Desalting
4:10
Oxygen Electrode Preparation
5:41
Oxygen Electrode Kinetic Assay
6:48
Results: Representative DesB Dioxygenase Induction, Activity, and Inhibition
7:50
Conclusion
Transcript
This method is designed to answer key questions in the field inorganic biochemistry, about how to maintain enzyme activity in proteins that contain oxygen-sensitive metals. The main advantage of this technique is that it helps you visualize ways i
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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.