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Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
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

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10:00 min

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November 08, 2016

DOI:

10.3791/54569-v

DOI:
10.3791/54569-v

•

10:00 min
November 08, 2016

•

2 Views
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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

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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.

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

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Villanueva-Cabello, T. M., Martinez-Duncker, I. Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy. J. Vis. Exp. (117), e54569, doi:10.3791/54569 (2016).

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