流式细胞术
CD8型
生物
免疫染色
免疫印迹
肺
细胞生物学
免疫学
免疫系统
分子生物学
医学
免疫组织化学
遗传学
基因
内科学
作者
Kerstin Sauer,Petra Scholtes,Roman Karwot,Susetta Finotto
出处
期刊:Nature Protocols
[Springer Nature]
日期:2006-12-01
卷期号:1 (6): 2870-2875
被引量:68
标识
DOI:10.1038/nprot.2006.435
摘要
The regulation of the cellular immune response in lung diseases is not yet fully understood. Isolating different subsets of immune cells directly from the lung is therefore an indispensable method of gaining detailed knowledge on the function of these cells in this organ. This protocol describes a method of isolating and magnetically labeling CD4+ lung T cells, which are then loaded and retained on the column while all other cells run through it (positive selection). The yield of this isolation is approximately 5 x 10(5) to 1.5 x 10(6) CD4+ cells from a murine lung. These cells can be further investigated by several methods such as flow cytometry, western blot analysis, RT-PCR, immunostaining and ELISA. In addition, lung CD4+ T cells alone or along with other immunologically important cells such as CD8+ T cells and T regulatory cells can be adoptively transferred into immune-deficient mice, and can influence important local parameters. This protocol can be completed in approximately 4 h 20 min.
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