BIOL2144Cellular Communication Semester 1 2022
BIOL2144Cellular Communication Semester 1 2022
Practical 4 Report
Laboratory skills to study cell communication -
SDS-PAGE and Western blots
Assignment 10% of overall grade (report is out of 30 marks)
Name:
Student number:
Activity 3 Data Analysis to identify a mystery TRP channel
Questions to be completed in this report
Q1. Based on the electrophysiology experiment, what temperature range does your TRP channel respond to? Can you eliminate any channels, why or why not? Can you identify your TRP channel? Why or why not? (2 marks)
Q2. What does this calcium imaging experiment tell you about your TRP channel? Please explain. Can you eliminate any channels, why or why not? Can you identify your TRP channel? Why or why not? (2 marks)
Q3. Explain the results from the animal behavior experiment and what you can conclude so far. Can you eliminate any channels, why or why not? Can you identify your TRP channel? Why or why not? (2 marks)
Q4. Explain the results from the antagonist experiment and what you can conclude so far. Can you eliminate any channels, why or why not? Can you identify your TRP channel? Why or why not? (2 marks)
Q5. What additional experiments could you do to confirm the function of your mystery TRP channel? (2 marks)
Q6. What chemical ligand, if any, activates your TRP receptor? How do you know this? (3 marks)
Q7. Bringing together all of the data, what TRP channel do you think you have and why? How confident are you in your conclusion? (6 marks)
Q8 Results BCA assay (5 marks)
Include your plate reader results and indicate your unknown data.
Plot the standard curve label this Figure 1 in your report. (1 mark)
Comment on the results that you have obtained. (1 mark)
From your graph and the measured absorbance values for your unknown sample, determine the concentration of protein in your sample and show how you determined and calculated the concentration of your cell lysate. (2 marks)
How much protein was in the sample you loaded onto the class SDS-PAGE gel? (1 mark)
Q9 SDS-PAGE theory questions (6 marks)
In the practical you separated proteins with polyacrylamide gel electrophoresis (PAGE) using MES running buffer that contained sodium dodecyl sulfate (SDS). What is the role of SDS?
Explain the principle of electrophoretic separation of the proteins contained in the lysates of human vascular smooth muscle cells by SDS-PAGE.
The gel cassettes you used in the prac contained two different layers of acrylamide between glass plates. An upper stacking layer and a lower resolving layer. Explain how the different layers act in SDS-PAGE.
