Preparation of biologically active subcellular fractions using the Balch homogenizer.
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.
German
Christopher L
CL
Program in Molecular Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA.
Howe
Charles L
CL
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
D002021:Buffers