Isolation and Purification of B Cells from Human Peripheral Blood
Published: July 31, 2023
Abstract
Source: Tzeng, S. J. The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity. J. Vis. Exp. (2016)
In this video, we demonstrate the isolation of B cells from human peripheral blood using biotinylated antibodies specific for non-B cells and streptavidin-conjugated magnetic microbeads.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board 1. Isolation and Purification of Human Peripheral Blood B Cells Draw ~ 10 mL of blood from the median cubital vein (in the cubital fossa anterior to the elbow) into a 15-mL tube containing K2EDTA (1.5 to 2.0 mg/mL blood) and immediately…
Disclosures
The authors have nothing to disclose.
Materials
| BD Vacutainer K2E | BD Biosciences | 367525 | 10 ml tube |
| Ficoll-Paque Plus | GE Healthcare | 17-1440-02 | Endotoxin-free |
| Trypan blue 0.5% solution | Biological Industries | 03-102-1B | Dye |
| IMag Human B lymphocyte enrichment set | BD Biosciences | 558007 | Reagent |
| Streptavidin magnetic microbeads | BD Biosciences | 9000810 | Reagent |
| 15 ml Falcon tubes | BD Falcon | 352196 | Plastic ware |
| Biotinylated CD27 mAb | Biolegend | 302804 | Clone O323 |
| Biotinylated anti-human CD27 | Biolegend | 302804 | Clone O323 |
| Biotinylated anti-human CD27 | eBioscience | 13-0279-80 | Clone O323 |
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Cite This Article
Isolation and Purification of B Cells from Human Peripheral Blood. J. Vis. Exp. (Pending Publication), e21523, doi: (2023).