脱颗粒
Percoll公司
免疫磁选
流式细胞术
差速离心
嗜酸性粒细胞
外周血
全血
分离(微生物学)
离心
免疫学
体外
CD16
化学
生物
色谱法
分子生物学
抗原
生物化学
生物信息学
CD8型
受体
CD3型
哮喘
作者
Kiho Son,Paige Lacy,Manali Mukherjee
出处
期刊:Methods in molecular biology
日期:2021-01-01
卷期号:: 15-25
被引量:1
标识
DOI:10.1007/978-1-0716-1095-4_2
摘要
The choice of isolation technique for human peripheral blood eosinophils contributes to the understanding of clinically relevant data derived from in vitro research. Since the 1990s, eosinophils have been conventionally isolated via density gradient centrifugation followed by negative immunomagnetic selection using anti-CD16 antibody-coated magnetic beads. Due to recent advancements in molecular techniques, “newer” methods have been made commercially available that drastically reduce user handling and processing time while maintaining high population purity. Here, we describe an isolation procedure using one of these methods, the human MACSxpress® Whole Blood Isolation Kit, as well as outline protocols for differential staining and flow cytometry analysis to evaluate the purity and activation state of isolated cells. In addition, we highlight an in vitro degranulation assay that may be used to verify the intact functionality of the isolated eosinophils.
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