Use chromatography to purify a protein of interest.
Use a colorimetric assays to examine protein activity and folding.
Determine the inhibition of several antibiotics with the protein.
Determine how various reagents alter the stability/activity of the protein.
Examine protein purity using gel electrophoresis.
Generate a protein concentration standard curve and calculate the concentration of your
protein solutions.
Use PyMol to visualize β-lactamase proteins.
What are the different methods used to purify proteins from crude extract materials? Why are
so many different types of methods required?
• What is happening in each step of our purification protocol?
• How does gel electrophoresis work? What characteristics of the protein determine how far it
will migrate in a gel? We will run denaturing gels, what does this mean and why is it
important?
• How do various reagents, such as urea, hydrogen peroxide and change in pH, affect protein
activity and stability?
• What is a protein’s “specific activity”?
• What is the basis for the nitrocefin-based activity assay? How does it work?
• How does Bis-ANS enable visualization of protein folding?