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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

Article DOI: 10.3791/54569-v • 10:00 min • November 8th, 2016
November 8th, 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

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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 Cells GD3 Ganglioside GD2 Ganglioside Microscopy Staining Cell Surface Mononuclear Cells Membrane Glycolipids Ganglioside Expression CD4+ T Cell Suspension Low Number Of Cells Sample Optimization T Cell Leukemia Peripheral Blood Sterile PBS EDTA 1.077 Density Sucrose Solution Centrifugation Peripheral Blood Mononuclear Cells Washing Naive CD4+ T Cells Negative Selection Magnetic Beads Purity CD4
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