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Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

DOI:
10.3791/53157-v

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19:05 min

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October 30, 2015

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, ,
1Institute for Applied Physics – Biophysics,Vienna University of Technology, 2Institute for Hygiene and Applied Immunology,Medical University of Vienna

Chapters

  • 00:00Title
  • 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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: TCR-pMHC BindingFRET-based MicroscopyT-cell Antigen SensitivityPlanar Lipid BilayerSingle Molecule FRETReceptor-ligand Interactions

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