Lab 9: Determination of Protein Stability using Chemical Denaturation and Intrinsic Fluorescence
Lab 9 Objectives
- Measure the free energy of unfolding (ΔGunfolding) of wild type and mutant HCAII using intrinsic protein fluorescence
- Determine whether your mutation affects protein stability
NOTE: For this and all following labs you will be working as a group of four in your pods and you will need to develop much of the procedures yourselves. Try to find efficient ways to complete the experiments; i.e., don’t have one person pipette everything while three people sit around and watch. You will still turn in one lab report per pair, but each pair will be responsible for reporting on data for both the wt and mutant proteins.