Centelles J J, Franco R, Canela E I, Bozal J
Biol Chem Hoppe Seyler. 1986 Apr;367(4):307-12. doi: 10.1515/bchm3.1986.367.1.307.
The rat brains homogenized with different media (sucrose, ethylene glycol, dimethyl sulfoxide and urea) yielded different amounts of microsomal fractions. The dielectric constant, density and viscosity of the homogenization media did not correlate with the amount of microsomes separated by differential centrifugation. The homogenization media containing dimethyl sulfoxide were the most efficient for the isolation of rat brain microsomes. The increase in the yield was up to 4-fold when 50% (v/v) dimethyl sulfoxide was employed. Microsomes isolated in this manner were analogous to those obtained from isotonic sucrose solution, as was demonstrated by their chemical and enzymatic (5'-nucleotidase, adenosine deaminase, guanine deaminase, purine-nucleoside phosphorylase, lactate, malate and glutamate dehydrogenases, amine oxidase fumarate hydratase, acid and alkaline phosphatase, acetylcholinesterase, NADPH-cytochrome c reductase, catalase and thiamine-diphosphatase) characterization.
用不同介质(蔗糖、乙二醇、二甲基亚砜和尿素)匀浆的大鼠脑产生了不同量的微粒体部分。匀浆介质的介电常数、密度和粘度与通过差速离心分离的微粒体数量无关。含有二甲基亚砜的匀浆介质对大鼠脑微粒体的分离最为有效。当使用50%(v/v)二甲基亚砜时,产量增加高达4倍。以这种方式分离的微粒体与从等渗蔗糖溶液中获得的微粒体相似,这通过它们的化学和酶学(5'-核苷酸酶、腺苷脱氨酶、鸟嘌呤脱氨酶、嘌呤核苷磷酸化酶、乳酸、苹果酸和谷氨酸脱氢酶、胺氧化酶、延胡索酸水合酶、酸性和碱性磷酸酶、乙酰胆碱酯酶、NADPH-细胞色素c还原酶、过氧化氢酶和硫胺二磷酸酶)特性得到证明。