A student runs an enzyme assay using the anaerobic glycolytic enzyme, lactate dehydrogenase (LDH). Their 1 ml assay is: 900 μl 200 mM Tris, pH 7.335 μl 30 mM pyruvate (in H2O)35 μl 6.6 mM NADH (in H2O) 30 μl of their LDH solution, in 200 mM Tris, pH 7.3They mix everything gently together in a cuvette and then measure the ΔA340/Δtime using a spectrophotometer. Their measured ΔA340/30 sec. was “-0.0311”.question : d) Let’s say that the total time of your LDH assay was 2 minutes. Did the pH of the assay change much during that time? Show a calculation. (Model your calculation after questions #40 and #41 on page 93 of Biochemical Calculations). The pKaof Tris is 8.21. No, the pH remained at 7.3. Use two HendersonHasselbalch equations.

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