FACS Staining for Follicular Helper T Cells
滤泡性辅助性T细胞的流式细胞分析
发布: 2012年01月20日第2卷第2期 DOI: 10.21769/BioProtoc.35 浏览次数: 22860
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Abstract
Germinal center is the primary site for B cells to undergo somatic hypermutation and class switching. A recently discovered subset of T cells known as T follicular helper (TFH) cells are essential for germinal center reaction and thus have been the subjects of intensive study in the recent years. This protocol describes a reliable way to stain this population for flow analysis. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research.
Keywords: Flow cytometry (流式细胞仪)
Mouse (小鼠)
Follicular helper T cells (滤泡辅助性T细胞)
Germinal center (生发中心)
Background
Materials and Reagents
- Antibodies
- Rat anti-mouse CXCR5-biotin (BD Biosciences, Pharmingen™, catalog number: 551960 )
- Goat anti-rat IgG-biotin (BD Biosciences, Pharmingen™, catalog number: 554014 )
- Straptavidin-PE Cy7 (BD Biosciences, Pharmingen™, catalog number: 557598 )
- Rat anti-mouse B220-pacific blue 450 (eBioscience, catalog number: 48-0452-82 )
- Rat anti-mouse CD11b-pacific blue 450 (eBioscience, catalog number: 48-0112-82 )
- Rat anti-mouse CD4-APC eFluor 780 (eBioscience, catalog number: 47-0042-82 )
- Hamster anti-mouse PD-1-PE (eBioscience, catalog number: 12-9985-82 )
- Mouse Fc block (BD Biosciences, Pharmingen™, catalog number: 553141 )
- Rat anti-mouse CXCR5-biotin (BD Biosciences, Pharmingen™, catalog number: 551960 )
- Other materials
- Single cell suspensions derived from murine spleen samples
- Fetal bovine serum (FBS) (Hyclone)
- Dulbecco’s modification eagle medium (DMEM) (Life Technologies, Invitrogen™)
- Ammonium chloride (0.17 M, filtered, autoclaved) (pH7.4)
- DAPI nucleic acid stain (Life Technologies, Invitrogen™)
- FACS buffer (see Recipes)
- Single cell suspensions derived from murine spleen samples
Equipment
- BD LSR II flow cytometer
- Conical tubes (Corning)
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Liu, Z. (2012). FACS Staining for Follicular Helper T Cells. Bio-protocol 2(2): e35. DOI: 10.21769/BioProtoc.35.
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分类
免疫学 > 免疫细胞染色 > 流式细胞术
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