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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
 

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

Article DOI: 10.3791/54582-v • 08:26 min • December 14th, 2016
December 14th, 2016 •
1
1Graduate Institute of Pharmacology, College of Medicine, National Taiwan University

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Summary
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

ELISpot Human Antibody-secreting B Cells Blood Quantification Isolation Differentiation Functional Readout Humoral Immunity Biomedical Field Immunology Vaccine Research Single Cell Level Tsung-Chih Tseng Graduate Student Laboratory Blood Sample Median Cubital Vein Potassium Supplemented EDTA Red Blood Cell Lysis Buffer Incubation Centrifuge White Pellet Residual Lysis Buffer Autoclaved PBS RPMI 1640 Medium Culture Dish 37 Degrees Celsius Five Percent Carbon Dioxide
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