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Chapters
- 00:00Title
- 00:29Preparing the lipids
- 02:13Determining how much ICAM-1 and MHC are deposited on the planar bilayer
- 03:22Cleaning the glass cover slips for forming planar bilayers
- 04:47Forming the bilayers
- 08:27Charging the bilayers and introducing His-tagged proteins
- 10:20Introducing primary T cells into the flow cell
- 11:36Conclusion
Supported planar bilayers are powerful tools that can be used to model the molecular interactions in an immunological synapse. Here, we show methods for anchoring cell adhesion proteins known to modulate synapse formation to the upper leaflet of the lipid bilyer and visualize synapse formation using TIRF microscopy.
Tags
Supported Planar BilayersImmunological SynapseKinapseMolecular InteractionsLymphocytesAntigen Presenting CellsNi2+-chelating LipidsRecombinant Cell Adhesion ProteinsMHC ProteinsPoly-histidine TagsGlass SurfacesSpecialized ChamberFlow-cellTIRF MicroscopySynapse FormationT Cell LocomotionReceptor SortingReceptor Degradation
