小中大
最近又有战友发过类似问题, 如下方法:
The membrane and cytosolic fractions were prepared as described (Souza & Ramirez, 1991) with minor modifications. Briefly, cerebral cortex was homogenized in 5 volumes of ice-cold homogenizing buffer containing 20 mM Tris–HCl buffer (pH 7.4), 1 mM DTT, 5 mM EGTA, 2 mM EDTA, 10% glycerol, 1 mM MgCl2, 1 mM PMSF, 2 μg/ml of leupeptin, and 2 μg/ml of aprotinin by dounce homogenizer with the use of pestle ‘B’ (Wheaton Scientific, Millville, NJ, USA), which allows disruption of tissue without disrupting the nuclei. The homogenate was centrifuged twice at 1,000 g for 15 min, and the pellet was discarded. The supernatant was further centrifuged at 27,000 g for 1 h at 4°C, and the supernatant was designated as the crude cytosolic extract. The pellet was resuspended in homogenizing buffer containing 0.5% Triton X-100, incubated for 1 h at 4°C, and centrifuged at 27,000 g for 30 min. The supernatant obtained from this centrifugation was designated as enriched membrane fraction. Protein content was assayed by the method of Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, pp. 248–254. Abstract | View Record in Scopus | Cited By in Scopus (75143)Bradford (1976).
源文献：
Effect of prenatal and postnatal ethanol exposure on Ca2+ /calmodulin-dependent protein kinase II in rat cerebral cortex.Mahadev K, Chetty CS, Vemuri MC.Alcohol. 2001 Apr;23(3):183-8
以上贴出自: cuturl('http://www.dxy.cn/bbs/post/view?bid=65&id=14454881&sty=1&tpg=1&age=0')
可以试试看, 至少有了参考文献. 呵呵.
