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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