Trizol法提取大鼠耳蜗髓磷脂P0蛋白质

The investigation of the method of extracting the rat's cochlea myelin protein zero with Trizol

目的用Trizol法提取大鼠耳蜗髓磷脂P0蛋白质(myelin protein P0,P0蛋白),与传统的蛋白裂解液裂解法比较蛋白提取效果,分析Trizol法的优势。方法Trizol法和传统的蛋白裂解液裂解法分别提取200g～250g Wistar大鼠(雌雄不限)耳蜗P0蛋白,用WesternBlot法及考马斯亮兰(Bradford)法比较两种方法提取P0蛋白含量及效果。结果Trizol法提取的大鼠耳蜗P0蛋白与传统的蛋白裂解液裂解法提取的含量相当,效果肯定。结论Trizol法与传统的蛋白裂解液裂解法提取的蛋白效果相当,但Trizol法简化了操作步骤,节省了单独提取蛋白质所需的蛋白酶抑制剂,节约样本,节省时间,提高工作效率,减少了蛋白和核酸降解的机会,有着更好的前景。

OBJECTIVE To extract rat＇s cochlea myelin protein PO by Trizol and compared with the protein extracting method of decomposition fluid in order to find the advantage of Trizol method. METHODS Cochlea myelin protein PO of the Wistar rat （200g-250g） was extracted by Trizol and the traditional protein decomposition fluid crack solution separately, authenticated by Western Blotting and Bradford. RESULTS The content and effect of two methods of extracting rat＇s cochlea myelin protein PO are similar. CONCLUSION The effect of two methods of extracting the protein was similar, but the method of Trizol had more advantages. Such as it can simplify the sequence of operation , save the time, enhance the working efficiency, moreover reduce the possibility of protein and RNAdegradation. So the Trizol method has the better prospect.