Peer-reviewed
Subcellular Fractionation Using Accudenz Gradient to Separate ER/Golgi in Yeast
使用Accudenz细胞组分梯度离心法分离酵母中的ER/Golgi
发布: 2012年01月20日第2卷第2期 DOI: 10.21769/BioProtoc.20 浏览次数: 16300
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Abstract
This protocol describes how to separate the endoplasmic reticulum (ER) and Golgi apparatus in yeast cells using a subcellular fractionation approach with an Accudenz gradient.
Materials and Reagents
- Accudenz (Accurate Chemical & Scientific Corporation)
- Protease inhibitors
- Phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich, catalog number: 78830-5G )
- Aprotinin (Sigma-Aldrich, catalog number: A3428-10MG )
- Pepstatin A (Sigma-Aldrich, catalog number: P5318-5MG )
- Phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich, catalog number: 78830-5G )
- NaN3
- Sodium fluoride (NaF)
- Tris-HCl
- Beta-mercaptoethanol
- TCA
- Sorbitol
- HEPES-KOH
- Spheroplasting buffer (see Recipes)
- Lysis buffer (see Recipes)
Equipment
- Dounce homogenizer (Cole-Parmer)
- Refractometer (Bausch & Lomb Incorporated)
- Beckman rotor
Procedure
English
中文翻译
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Editor, B. (2012). Subcellular Fractionation Using Accudenz Gradient to Separate ER/Golgi in Yeast. Bio-protocol 2(2): e20. DOI: 10.21769/BioProtoc.20.
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分类
微生物学 > 微生物细胞生物学 > 细胞器分离
细胞生物学 > 细胞器分离 > 高尔基体
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