Purpose

In this experiment, you will collect data that will allow you to determine whether the kinetic mechanism of your inhibitor is competitive, uncompetitive, non-competitive, or mixed. Just like last week, you will substrate concentrations will be varied to determine the kinetic parameters V_max/K_m, V_max, and K_m for your SDS-activated enzyme sample. However, this week, an inhibitor will be present.

Objectives

Upon successful completion of this lab, you will be able to:

• Design and execute protocols for estimating inhibition kinetics parameters K_i.

• Explain the relationship difference between K_i and IC₅₀.

• Determine the kinetic mechanism of an inhibitor by comparing the change in the kinetics parameters V_max, V_max/K_m, and K_m using Eadie-Hofstee plots.

• Write a complete Introduction in the style of a professional research report.

 

Pre-lab Preparation

Read the assigned reading (including this lesson) completely

Complete the prelab assignment at least 24 hours before lab begins

Outline the purpose, objectives, and procedures sections in your notebook

 Lecture Video

[Lecture Slides]