Microsome Isolation from Tissue
从组织中分离微粒体
发布: 2014年02月05日第4卷第3期 DOI: 10.21769/BioProtoc.1038 浏览次数: 12920
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参见作者原研究论文
The authors used this protocol in:
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
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:
- Bodero, M. and Abisambra, J. F. (2014). Microsome Isolation from Tissue. Bio-protocol 4(3): e1038. DOI: 10.21769/BioProtoc.1038.
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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.
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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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