晶状体蛋白质双向凝胶电泳的可重复性及分辨率研究

Research on repeatability and resolution of lens two-dimensional electrophoresis

目的 对晶状体蛋白质双向凝胶电泳进行可重复性及分辨率研究。方法 用固相pH梯度等电聚焦 (IPG DALT)的方法 ,分别选用不同的上样量 ,10 %或 13 %的SDS 聚丙烯酰胺凝胶电泳 ,用银染或考马斯亮蓝染色的方法对 3个月新西兰兔的晶状体蛋白进行分离。结果 当上样量为 15 0 μg ,13 %的分离胶对晶状体蛋白 (crystallin)的分离效果较好 ,当上样量为 1 2mg ,10 %的分离胶对高相对分子质量蛋白的分离效果较好。凝胶点的匹配率为 83 5 %和 91 7%。 结论 用IPG DALT的方法 ,综合小样本量和低密度凝胶电泳与大样本量高密度凝胶电泳结果 ,可以更好地分析晶状体蛋白的分离效果 。

Objects To study the repeatability and resolution of lens protein two-dimensional electrophoresis(2-DE). Methods Lens proteins form three-month-old rabbit were separated using 2-DE using series SDS-PAGE density (10% or 13%) and stain methods. Results Crystallin could be well separated from 150 μg of total protein sample using 13% SDS-PAGE.High molecular weight proteins could be well separated from 1.2 mg sample using 10% SDS-PAGE.The matching rates were 83.5%91.7%. Conclusion Using IPG-DALT method,electrophoresis provide better analysis for the lens protein separation,but the data repeatability should be further improved.