Propidium Iodide Staining of Cells for FACS Analysis
流式细胞术检测细胞增殖与凋亡（碘化丙啶法）
发布: 2012年06月05日第2卷第11期 DOI: 10.21769/BioProtoc.195 浏览次数: 58371
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Abstract
Fluorescence Activated Cell Sorting (FACS) is used to study DNA cell content. Propidium iodide (PI) intercalates into double-stranded nucleic acids and fluoresces. It is excluded by viable cells but can penetrate cell membranes of dying or dead cells. Thus PI staining is included in immunofluorescent staining protocols to identify dead cells. DNA staining can be used to study the cell cycle. Relative DNA content shows the proportion of cells in G1, G2 and S phases. Apoptotic cells show characteristic smear on DNA staining. Here a protocol to stain cells by PI is described.
Keywords: Propidium iodide (碘化丙啶)
FACS (流式细胞仪)
Cell cycle (细胞周期)
Materials and Reagents
- Triton X-100 (Sigma-Aldrich, catalog number: T9284 )
- Propidium iodide (PI) (Sigma-Aldrich, catalog number: P4170 )
- RNase A (Sigma-Aldrich, catalog number: R4642 )
- Phosphate buffered saline (PBS)
- 70% EtOH
- 10 % Triton X-100 (see Recipes)
- PI stock solution (see Recipes)
Equipment
- Standard table-top centrifuges
- FACS machine
- Incubator
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Zhu, H. (2012). Propidium Iodide Staining of Cells for FACS Analysis. Bio-protocol 2(11): e195. DOI: 10.21769/BioProtoc.195.
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分类
癌症生物学 > 增殖信号转导 > 细胞生物学试验 > 细胞周期
癌症生物学 > 通用技术 > 细胞生物学试验 > 细胞周期
细胞生物学 > 细胞染色 > 核酸
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