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 language | English (US) |
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Pages (from-to) | 117-124 |
Number of pages | 8 |
Journal | Analytical Biochemistry |
Volume | 394 |
Issue number | 1 |
DOIs | |
State | Published - Nov 1 2009 |
Keywords
- Balch homogenizer
- Centrifugation
- Endosome
- IL-6
- STAT
- Vesicle isolation
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology