Dissipative Microgravimetry Technique to Study Protein-Lipid Bilayer Interaction
Transcript
Carefully dock the plasma-cleaned sensors into the four flow chambers using tweezers. Avoid any pressure on or torsion of, the chambers and tubes that may cause leaking. Flush the system with citrate buffer in the open flow mode for 10 minutes.
Launch the program. Start recording any changes in the frequency and dissipation of the first fundamental tone and overtones using the software until the frequency and dissipation baselines are stable.
When the baselines are stable, apply the SUV suspension in citrate buffer. Using a reaction vessel, remove 1.5 milliliters of the dead volume. Then, close the system in the loop flow mode. Record the frequency dissipation shift for another 10 minutes.
When the SLB is stable, equilibrate the system with the running buffer at the required calcium concentrations in an open-flow mode for 40 minutes. Add the protein to the running buffer containing calcium. Perform the application of the protein in a loop-flow mode until an equilibrium steady state is reached.
