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The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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Immunology and Infection
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JoVE Journal Immunology and Infection
The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

DOI:
10.3791/54582-v

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08:26 min

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December 14, 2016

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1Graduate Institute of Pharmacology, College of Medicine,National Taiwan University

Chapters

  • 00:05Title
  • 00:48Human Peripheral Blood B Cell Isolation and Purification
  • 02:57Naïve and Memory B Cell and Plasmablast/Plasma Cell Sorting
  • 03:54In Vitro Isolated Human CD19+, CD19+CD27+ Memory, and CD19+CD27– Naïve B Cell Differentiation and ELISpot Analysis
  • 06:18Results: Representative ELISpot Analysis
  • 07:06Conclusion

Summary

Automatic Translation

Automatic Translation

Human peripheral blood is commonly used for the assessment of the humoral immune response. Here, the methods for isolating human B cells from peripheral blood, differentiating human B cells into antibody (Ab)-secreting B cells (ASCs) in culture, and enumerating the total IgM- and IgG-ASCs via an ELISpot assay are described.

Tags

Keywords: ELISpotAntibody-secreting B CellsBloodHumoral ImmunityQuantificationDifferentiationIsolationVaccine ResearchImmunologySingle Cell DetectionBiotinylated AntibodyStreptavidin MicrobeadsMagnetic Separation

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