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Microsome Isolation from Tissue

从组织中分离微粒体

MB Maria Bodero
Jose Francisco Abisambra Jose Francisco Abisambra

发布: 2014年02月05日第4卷第3期 DOI: 10.21769/BioProtoc.1038 浏览次数: 12979

评审: Hong-guang XiaXuecai Ge

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The authors used this protocol in:

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Abstract

This protocol details the extraction of microsomes from frozen tissue in order to further examine the protein-protein interactions occurring within the endoplasmic reticulum. This protocol was adapted from Abisambra et al. (2013) with modifications made in order to optimize for subsequent use.

Keywords: Endoplasmic reticulum (内质网) search Microsomes (微粒体) search Sub-cellular fractionation (亚细胞分馏) search

Materials and Reagents

  1. Sucrose
  2. Protease Inhibitor cocktail, EDTA free (Merck KGaA, Calbiochem, catalog number: 539134 )
  3. Phosphatase inhibitor cocktail II
  4. Phosphatase inhibitor cocktail III
  5. PMSF at 10 mM in DMSO or 1.74 mg/ml (Thermo Fisher Scientific, catalog number: 36978 )
  6. Phosphatase Arrest II cocktail (Geno Technology, catalog number: 786-451 )
  7. Phosphatase Arrest III cocktail (Geno Technology, catalog number: 786-452 )
  8. M-PER Mammalian Protein Extraction Reagent (Thermo Fisher Scientific, catalog number: 78501 )

Equipment

  1. Sterile bottle filter
  2. Glass Dounce homogenizer
  3. Refrigerated centrifuge
  4. Microfuge tubes rated for at least 25,000 x g centrifugation

Procedure

English
中文翻译

文章信息

版权信息

© 2014 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

  1. Bodero, M. and Abisambra, J. F. (2014). Microsome Isolation from Tissue. Bio-protocol 4(3): e1038. DOI: 10.21769/BioProtoc.1038.
  2. Abisambra, J. F., Jinwal, U. K., Blair, L. J., O'Leary, J. C., 3rd, Li, Q., Brady, S., Wang, L., Guidi, C. E., Zhang, B., Nordhues, B. A., Cockman, M., Suntharalingham, A., Li, P., Jin, Y., Atkins, C. A. and Dickey, C. A. (2013). Tau accumulation activates the unfolded protein response by impairing endoplasmic reticulum-associated degradation. J Neurosci 33(22): 9498-9507.

    3. ris ris Download Citation in RIS Format

分类

细胞生物学 > 细胞器分离 > 微粒体

生物化学 > 蛋白质 > 相互作用 > 蛋白质-蛋白质相互作用

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