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为建立适合于大葱花蕾总蛋白的双向电泳体系,本研究以大葱花蕾为材料,对大葱花蕾总蛋白提取方法、IPG胶条梯度范围、上样量、等电聚焦等双向电泳的关键步骤进行优化。结果表明最适宜大葱花蕾总蛋白的双向电泳条件为:TCA-丙酮沉淀法提取大葱花蕾总蛋白,选择24 cm p H 4~7的IPG胶条,上样量800μg,按聚焦程序Ⅰ聚焦后进行双向电泳并采用考马斯亮蓝染色。采用该体系可以获得分辨率高、重复性好的双向电泳图谱。
In order to establish a two-dimensional electrophoresis system suitable for the total protein of Chinese onion flower buds, this study optimized the key steps of two-dimensional gel electrophoresis, such as the method of extracting total protein from Chinese onion scallion bud, the gradient range of IPG gel strips, loading amount and isoelectric focusing . The results showed that the most suitable two-dimensional gel electrophoresis conditions were as follows: TCA-acetone extraction of Chinese onion flower buds total protein, select 24 cm p H 4 ~ 7 IPG strips, the amount of 800 μg, according to the focusing program Ⅰ focus Two-dimensional electrophoresis and staining with Coomassie brilliant blue. The system can be obtained with high resolution, good reproducibility of two-dimensional electrophoresis.