Immunofluorescence Analysis of Yeast Protein
酵母蛋白的免疫荧光分析
发布: 2012年07月05日第2卷第13期 DOI: 10.21769/BioProtoc.211 浏览次数: 17001
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Abstract
Many important regulatory proteins such as transcription factors are regulated through subcellular localization. Protein localization can be examined by fusing a GFP tag. However, GFP is relatively big in size, and potentially may affect correct protein localization. Several small tags have been developed, such as myc, HA or Flag. By using immunostain and fluorescence microscopy as described in this protocol, one can easily probe the regulation of a selected yeast protein with the application of the aforementioned small tags.
Keywords: Yeast (酵母)
Immunofluorescence (免疫荧光)
Microscope (显微镜)
Spheroplast (原生质体)
Antibody (抗体)
Materials and Reagents
- Yeast cells
- Potassium phosphate monobasic (KH2PO4) (Sigma-Aldrich, catalog number: P8416 )
- Potassium phosphate dibasic (K2HPO4·3H2O) (Sigma-Aldrich, catalog number: P9666 )
- Sorbitol (C6H14O6) (Sigma-Aldrich, catalog number: S1876 )
- BSA (Albumin from bovine serum) (Sigma-Aldrich, catalog number: A4503 )
- Potassium chloride (KCl) (Thermo Fisher Scientific, catalog number: BP366-1 )
- 37% formaldehyde solution (Thermo Fisher Scientific, catalog number: F75P1GAL )
- Zymolyase (USB, catalog number: Z1001 )
- Vectorshield
- Poly-L-lysine
- DAPI
- Cytoseal 60
- Phosphate buffer (see Recipes)
- Sorbitol buffer (see Recipes)
- Blocking buffer (see Recipes)
Equipment
- Centrifuges
- Shaker
- Conical tube
- Fluorescence microscope
- 15 ml conical tube
- Light microscope
- Heat blot
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Wei, Y. (2012). Immunofluorescence Analysis of Yeast Protein. Bio-protocol 2(13): e211. DOI: 10.21769/BioProtoc.211.
Download Citation in RIS Format
分类
微生物学 > 微生物生物化学 > 蛋白质
生物化学 > 蛋白质 > 荧光
生物化学 > 蛋白质 > 免疫检测
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