Lab 11: LAB ASSIGNMENTS
PRELAB QUIZ
Question 1
What experiment with HCAII are we simulating this week? What is the purpose of that experiment? (Provide 2-3 sentences).
Question 2
We will be studying acetazolamide (AZ) in this lab. What is AZ and why is it interesting? What medical applications does it have? (This will require a bit of research.)
Question 3
Use the BRENDA database again to find published Ki values for acetazolamide inhibition of HCAII. This should give you a general idea for how strongly AZ binds HCAII. Which of the following is the closest approximation? (Note that in lab 11 you are actually measuring KD, not Ki. However, the Ki should still give you an idea of the strength of binding.) Here is the link to BRENDA (link opens in a new tab): http://www.brenda-enzymes.info/
- 5 nM
- 12 nM
- 12 mM
IN-LAB ACTIVITIES
- After joining BBCU, you and your partner will join a breakout room.
- You will use the in-lab time to work with your partner to complete the planning template file for this experiment.
- Start with the pilot for DNSA. Once you have this ready, a TA will share and discuss some data with you.
- Next, plan the full DNSA experiment.
- The next step is to plan the pilot for AZ. A TA will take a look at your suggested high and low AZ concentrations and give you estimated AZ values. Based on these, you will likely need to adjust your AZ concentrations and try again.
- Once you and your TA are satisfied with your AZ concentrations, you can complete the planning document by filling out the full AZ experiment.
The following videos were made to help you understand how to plan these experiments, which are quite complicated:
Part 1: DNSA
TA video will be added when available
Part 2: AZ
TA video will be added when available
- You can use the remaining lab time with your lab partner to analyze the lab 11 stock data.
- TAs can be available for questions throughout lab. This lab requires several check-ins with TAs, so be aware that you may have to wait. (This is a good simulation of the live version of the lab, where there is always a line to use the plate reader.)
- Complete the lab/partner/self evaluation Google form.
- Get a sharable link to your Google sheets planning document and submit it to Canvas.