title: Preparation of biologically active subcellular fractions using the Balch homogenizer.
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.
authors.affiliation: Program in Molecular Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: German
authors.forename: Charles L
authors.initials: CL
authors.lastname: Howe
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002021:Buffers; D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism; D002498:Centrifugation; D003593:Cytoplasm / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D051379:Mice; D015203:Reproducibility of Results; D013347:Subcellular Fractions / Q000378:metabolism*; D013997:Time Factors
chemical_list: D002021:Buffers