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Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy
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Immunology and Infection
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JoVE Journal Immunology and Infection
Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy

Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy

DOI:
10.3791/54569-v

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10:00 min

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November 08, 2016

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1Instituto de Investigación en Ciencias Básicas y Aplicadas, Centro de Investigación en Dinámica Celular, Laboratorio de Glicobiología Humana y Diagnóstico Molecular,Universidad Autónoma del Estado de Morelos

Chapters

  • 00:05Title
  • 01:00Isolation and Activation of Human Naïve CD4+ T Cells
  • 04:22Collection and Fixation of Resting and Activated Naïve CD4+ T Cells
  • 05:34Staining, Mounting and Visualization by Confocal Microscopy
  • 07:47Results: Localization of GD3 and GD2 Gangliosides in Adhered Activated CD4+ T Cells
  • 09:13Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a standard antibody staining protocol for use in microscopy to determine the membrane expression and localization of gangliosides in resting and activated human naïve CD4+ T cells. Also described are real-time PCR experiments using <40,000 cells that do not require additional low input RNA kits.

Tags

Keywords: CD4+ T CellsGD3 GangliosideGD2 GangliosideMembrane ExpressionMicroscopyPeripheral Blood Mononuclear Cells (PBMCs)Negative SelectionAnti-CD3 AntibodyAnti-CD28 AntibodyCell StimulationCell CultureCell Preparation

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