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Anal Biochem. 2009 Nov 1;394(1):117-24. doi: 10.1016/j.ab.2009.07.024. Epub 2009 Jul 19.

Preparation of biologically active subcellular fractions using the Balch homogenizer.

Author information

1
Program in Molecular Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA.

Erratum in

  • Anal Biochem. 2010 May 15;400(2):310.

Abstract

Obtaining vesicular fractions from cell lines or animal tissue is both time and technically intensive. The presence of plasma membrane and nuclear contaminants within a preparation is often dependent on the method of homogenization and is usually mitigated through the use of density gradients. We have developed a method that utilizes Balch homogenization and differential centrifugation to obtain two distinct vesicular fractions along with purified nuclear, cytoplasmic, and ghost fractions within a 3-h period of time without the use of density gradients. Importantly, these fractions maintain their biologic activity following isolation and may be used for both localization and biochemical analyses.

PMID:
19622341
PMCID:
PMC2774742
DOI:
10.1016/j.ab.2009.07.024
[Indexed for MEDLINE]
Free PMC Article

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