从组织中分离微粒体

Maria Bodero; Jose  Francisco Abisambra

doi:10.21769/BioProtoc.1038

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Peer-reviewed

Microsome Isolation from Tissue

从组织中分离微粒体

MB Maria Bodero

Jose Francisco Abisambra email

发布: 2014年02月05日第4卷第3期 DOI: 10.21769/BioProtoc.1038 浏览次数: 12289

评审: Hong-guang XiaXuecai Ge

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参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Neuroscience, featuring study using the protocol.

May 2013

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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)

Microsomes (微粒体)

Sub-cellular fractionation (亚细胞分馏)

Materials and Reagents

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

Equipment

Sterile bottle filter
Glass Dounce homogenizer
Refrigerated centrifuge
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

如何引用

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

Bodero, M. and Abisambra, J. F. (2014). Microsome Isolation from Tissue. Bio-protocol 4(3): e1038. DOI: 10.21769/BioProtoc.1038.
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.