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Preparation of Bead-supported Lipid Bilayers to Study the Particulate Output of T Cell Immune Synapses
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Immunology and Infection
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JoVE Journal Immunology and Infection
Preparation of Bead-supported Lipid Bilayers to Study the Particulate Output of T Cell Immune Synapses

Preparation of Bead-supported Lipid Bilayers to Study the Particulate Output of T Cell Immune Synapses

DOI:
10.3791/63130-v

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11:06 min

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April 01, 2022

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1Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences,The University of Oxford

Chapters

  • 00:04Introduction
  • 01:07Protein Density Calibrations on BSLBs
  • 01:47Titration of His-Tagged Proteins on 12.5 mol% of DGS-(Ni) NTA-Containing BSLBs
  • 04:02Measurement of T-cell Output Transfer to BSLBs
  • 08:13Results: FCM for Absolute Protein Quantification on the Cell Surface
  • 10:17Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present the protocol for the stepwise reconstitution of synthetic antigen-presenting cells using Bead-Supported Lipid Bilayers and their use to interrogate the synaptic output from activated T cells.

Tags

Keywords: Bead-supported Lipid BilayersImmune SynapseFlow CytometryMicroscopyProteomicsRNA SequencingChimeric Antigen ReceptorsSignaling PathwaysGenetic AblationProtein CalibrationMulticolor Flow CytometrySilica BeadsLiposomeNickel-containing PhospholipidsArgonNitrogenBlockingBSANickel SulfateHBS-HSA BufferProtein Dilutions

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