Journal
ANALYTICAL BIOCHEMISTRY
Volume 394, Issue 1, Pages 117-124Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2009.07.024
Keywords
Balch homogenizer; Vesicle isolation; Centrifugation; Endosome; STAT; IL-6
Funding
- Mayo Clinic
- Mayo Graduate School
- Kern Predoctoral Neuroscience Fellowship
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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. (C) 2009 Elsevier Inc. All rights reserved.
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