Lab 4 results_ Enzyme Kinetic Analysis of Alkaline Phosphatase

Table 1: Substrate Hydrolysis analysis for Row A Well A1-A2 A3-A4 A5-A6 A7-A8 A9-A10 A11-A12 Substrate Concentra tion (mM) 0 0.05 0.1 0.5 1 2.5 Substrate Volume ( µ𝐿) 230 230 230 230 230 230 Water ( µ𝐿) 20 20 20 20 20 20 Table 2: Substrate concentration Kinetics for Row B Well B1-B2 B3-B4 B5-B6 B7-B8 B9-B10 B11-B12 Substrate Concentra tion (mM) 0 0.05 0.1 0.5 1 2.5 Substrate Volume ( µ𝐿) 230 230 230 230 230 230 Water ( µ𝐿) 10 10 10 10 10 10 Enzyme ( µ𝐿) 10 10 10 10 10 10 Table 3: Inhibitor Kinetics for Row C Well C1-C2 C3-C4 C5-C6 C7-C8 C9-C10 C11-C12 Substrate Concentra tion (mM) 0 0.05 0.1 0.5 1 2.5 Substrate Volume ( µ𝐿) 230 230 230 230 230 230 Inhibitor 10 10 10 10 10 10

( µ𝐿) Enzyme ( µ𝐿) 10 10 10 10 10 10 After the wells were filled with the desired concentrations of solutions, the microplate was taken to the microplate reader and Row A was read. Afterwards, enzyme was added to Row B using a multi-channel pipette and the solutions were gently mixed using the tips of the pipette. The plate was then reinserted into the microplate reader to read row B. The process used for row B was repeated for row C and the data provided below was obtained. Results: Part 1: Figure 1. Graph of the expected (change in enzyme concentration)