一种超滤管电洗脱装置及应用研究

An Ultrafiltration Tube Electroelution Device and Its Application

目的介绍一种自制超滤管电洗脱装置的制作方法,并与其他洗脱方法进行比较。方法以超滤管离心管、滤网和海绵制作成超滤电洗脱装置。待分离的蛋白样品用非变性聚丙酰胺凝胶电泳分离后染色、切胶、碎胶后,分别用上述自制超滤电洗脱装置、被动洗脱法和半透膜电洗脱法回收凝胶中的蛋白样品,再通过超滤离心进行脱盐浓缩。最后用Bradford法测定样品浓度,并比较三种洗脱方法的洗脱效率。结果超滤管电洗脱、被动洗脱和半透膜电洗脱分成0.5μg和5μg两组,以三种电洗脱法的蛋白回收率依次为:0.5μg组82%、44%、46%;5μg组77%、58.6%、63.6%。结论用超滤管自制的电洗脱装置制作简单方便,其回收效率比被动扩散法和半透膜电洗脱法高,特别适用于小量蛋白样品的回收。

Purpose To present the facture of a self-made uhrafiltration tube electroelution device and compare with passive elution and semi-permeable membrane electroelution. Methods Fistly one uhrafihration tube electroelution device was configured with an ultrafiltration tube, filter and sponge. Then we isolated the target protein by non-denaturing polyacrylamide gel electrophoresis, cut individual gel strips, and electroeluted them by three eletroelution methods of uhrafiltration tube electroelution,passive elution and semi- permeable membrane electroelution. After that we enriched and desalted the sample by uhrafiltration. Finally, using Bradford method we detected the protein concentration and assessed the recovery efficiency of three electroelution methods. Results Three electroelution methods of uhrafihration, passive diffusion and semi- permeable membrane elution were used into two groups of 0.5μg and 5μg. The recovery rates in 0.5 μg group were 82% ,44% ,46% with three eleetroelution methods respectively. In 5μg group,the recovery rates were 77% , 58.6% , 65.6% （5μg）. Conclusion The uhrafiltration electroelution device could be used for the recovery of protein in gel. Compared with other methods, the recovery efficiency of electroelution device was altrafihration higher than passive elution and semi-permeable membrane electroelution.