Preparation of biologically active subcellular fractions using the Balch homogenizer

Christopher L. German, Charles L Howe

Research output: Contribution to journalArticle

8 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)117-124
Number of pages8
JournalAnalytical Biochemistry
Volume394
Issue number1
DOIs
StatePublished - Nov 1 2009

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Keywords

  • Balch homogenizer
  • Centrifugation
  • Endosome
  • IL-6
  • STAT
  • Vesicle isolation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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