This experiment is based on the "Guide to Cellular Experiments", translated by Huang Peitang et al.
Operation method
Determination of Km and Vmax
Materials and Instruments
Protein Substrate ATP Analytical Tubes
Move
1. Prepare three sets of analytical tubes containing different amounts of each protein substrate to maintain the ATP concentration at a saturating concentration.
2. Perform the kinase assay.
3. perform an experiment in which the saturating concentration of the protein is kept constant by varying the concentration of ATP.
4. once the Km and Vmax values for the protein substrate and ATP are known, use this data to design optimal experimental conditions for the enzyme/substrate pair.
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