Binding Fluorescence Continued
The goal of this experiment was to measure the binding fluorescence of a large unilamellar lipid vesicle/peptide solution in order to measure the K of the reaction.
Refer back to part one of this experiment.
- Preparation of the Lipid Vesicles
- Add 3mL of the buffer into the pre-made lipid vesicles
- Vortex for 1 hour
- Mass of lipids = 0.025j
- 760.10 g/mol
- Concentration = 9.39 mM
- Extrusion of the lipid solution (method used to get unilamellar vesicles)
- Dilute stock lipid to 5μL, 25μL, and 100μL
- Obtain a baseline with the following:
- sodium phosphate buffer
- peptide solution of all concentrations
- lipid solution of all concentrations
- 1:1 dilution of each concentration at 25 °C
- Total of 9 combinations of lipid-peptide solutions
- Solution of 5μL lipids with 2μL peptide, 5μL lipids with 10μL peptide solution, and 5μL lipids with 30μL peptide solution
- Solution of 25μL lipids with 2μL peptide, 25μL lipids with 10μL peptide solution, and 25μL lipids with 30μL peptide solution
- Solution of 100μL lipids with 2μL peptide, 100μL lipids with 10μL peptide solution, and 100μL lipids with 30μL peptide solution
Data & Results
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