A simple protocol for the subcellular fractionation of skeletal muscle cells and tissue

BMC Research Notes

Malcolm J Jackson²  Daniela Caporossi¹  Timothy Pearson²  Ivan Dimauro¹ 

[1] Department of Health Sciences, University of Rome “Foro Italico”, Piazza Lauro De Bosis 15, 00194, Rome, Italy;Department of Musculoskeletal Biology, Institute of Ageing & Chronic Disease, University of Liverpool, Daulby Street, Liverpool L69 3GA, United Kingdom

关键词: Western blotting;    Subcellular fractionation;    Skeletal muscle;   

Others  :  1165648
DOI  :  10.1186/1756-0500-5-513

 received in 2012-04-05, accepted in 2012-08-31,  发布年份 2012

【 摘 要 】

Background

We describe a method for subcellular fractionation of mouse skeletal muscle, myoblast and myotubes to obtain relatively pure fractions of nuclear, cytosolic and mitochondrial compartments. Fractionation allows the analysis of a protein of interest (or other cellular component) based on its subcellular compartmental distribution and can also generate molecular information about the state of a cell and/or tissue and how the distribution of a protein may differ between different cellular compartments, tissues or cell types, in response to treatments or ageing.

Findings

The described method was specifically developed for skeletal muscle and proliferating/differentiated muscle cells. The purity of the different fractions, representing the cytoplasmic, mitochondrial and nuclear subcellular compartments was validated by western blot analysis of “house-keeper” marker proteins specific for each cellular compartment.

Conclusion

This low cost method allowed the mitochondrial, cytoplasmic and nuclear subcellular compartments from the same starting muscle samples to be rapidly and simultaneously isolated with good purity and without the use of an ultracentrifuge. This method permits samples to be frozen at −80°C for future analysis and/or additional processing at a later date.

【 授权许可】

   
2012 Dimauro et al.; licensee BioMed Central Ltd.

【 图 表 】

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