German Christopher L, Howe Charles L
Program in Molecular Neuroscience, Mayo Clinic College of Medicine, 200 First ST SW, Rochester, MN 55905, USA.
Anal Biochem. 2009 Nov 1;394(1):117-24. doi: 10.1016/j.ab.2009.07.024. Epub 2009 Jul 19.
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.
从细胞系或动物组织中获取囊泡组分既耗时又技术要求高。制剂中质膜和核污染物的存在通常取决于匀浆方法，并且通常通过使用密度梯度来减轻。我们开发了一种方法，该方法利用巴尔奇匀浆法和差速离心法，在不使用密度梯度的情况下，在3小时内获得两个不同的囊泡组分以及纯化的核、细胞质和空壳组分。重要的是，这些组分在分离后保持其生物活性，可用于定位和生化分析。
