Journal
/
Inmunología y contagio
/
Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy
JoVE Revista
Inmunología y contagio
Se requiere una suscripción a JoVE para ver este contenido.  Inicie sesión o comience su prueba gratuita.
JoVE Revista Inmunología y contagio
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

•

10:00 min

•

November 08, 2016

•

,
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

Capítulos

  • 00:05Título
  • 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

Traducción Automática

Traducción Automática

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

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

Article

Embed

Añadir a la lista de reproducción

Usage Statistics

Videos relacionados

Preparation and Use of HIV-1 Infected Primary CD4+ T-Cells as Target Cells in Natural Killer Cell Cytotoxic Assays

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells

Adenoviral Transduction of Naive CD4 T Cells to Study Treg Differentiation

Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

In Situ Detection of Autoreactive CD4 T Cells in Brain and Heart Using Major Histocompatibility Complex Class II Dextramers

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions

Imaging CD4 T Cell Interstitial Migration in the Inflamed Dermis

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Read Article