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Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
Capítulos
- 00:00Título
- 00:56Introduction
- 03:00Protein Production
- 04:35Calcium Flux Measurement
- 06:27FRET Donor Recovery after Acceptor Photobleaching
- 11:07Single Molecule FRET
- 16:29Determination of the Constant ‘C’
- 18:17Conclusion
This manuscript describes how to conduct (single molecule) Förster Resonance Energy Transfer (FRET)- based assays to measure the binding dynamics between T-cell antigen receptor (TCR) and antigenic peptide-loaded MHC molecules as they occur within the immunological synapse of a T-cell in contact with a functionalized planar supported lipid bilayer.
