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Preparation of biologically active subcellular fractions using the Balch homogenizer |
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| Authors: | Christopher L. German Charles L. Howe |
| Affiliation: | a Program in Molecular Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA b Department of Immunology, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA c Department of Neurology, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA d Department of Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA e Program in Translational Immunovirology and Biodefense, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA |
| 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. |
| Keywords: | Balch homogenizer Vesicle isolation Centrifugation Endosome STAT IL-6 |
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