Kinga — State the purpose of the entire experiment, delineating the methods that will be used to execute the experiment. Refer to the starting material as well as what the output of this experiment will be. For this offering, focus on the purification of three proteins- HaloTag-CLoG1, His-PPX, and GST-PPX substrate- and include how they will be used in concert in our purification project (as described in the Week 6 lectures).
Richard — Your group will be assigned one of the above three proteins( His-PPX this is the protein of interest you should focus) . Present a background on the protein itself (what it is, its size, properties, etc. are), and what it does biologically.
Serena — Based on the affinity tag that is already added to your protein, provide background on that affinity method. Include the general concept and theory behind how that tag and the resin are used to purify proteins, and state expected yields and purity based on the resin we will be using (product information posted on Canvas site).
Tara– Explain the key assessments for protein purification efficacy. You should focus on yield/recovery, purity/specific activity, and present quantitative and qualitative assessments you will employ (i.e., enzyme assay, protein assay, SDS-PAGE, immunoblot). Note: we will discuss these analytical procedures more in later weeks- do your best based on what we have talked about in lecture to date, and you can refine this section later.