The concentration of pyruvate in the cuvette.

Q4: In the practical you will determine the Km for pyruvate with LDH at a fixed concentration of NADH. To do this you will need to vary the concentration of pyruvate in the cuvette. The assay mix will be: 2.8 ml of assay buffer, 100 mM sodium phosphate pH 7.4 0.1 ml of sodium pyruvate 0.1 ml of 8 mM NADH To vary (decrease) the concentration of pyruvate In the cuvette you will keep the addition volume constant at 0.1 mL but decrease the concentration of the ovruvate stock solution that you add. The required final concentrations of pyruvate in the assay cuvette are listed in Table 2 in the manual (reproduced here). Complete Table 2 to show how you will prepare the stock solutions. Enter number only into the electronic test. Stock [Pyruvate] re • uired Volume and stock [Pyruvate] needed Volume of Final water for [Pyruvate] in dilution assa 24 mM 0.5 mL of 48 mM 0.5 mL Q4a 0.5 mL of 0.5 mL Q4b 0.5 mL of 0.5 mL Q4c 0.5 mL of 0.5 mL Table 2: Serial dilution of sodium pyruvate stock solutions 0.8 mM 0.4 mM 0.2 mM 0.1 mM