摘要
目的:研究人先天性核性白内障混浊晶状体蛋白质双向凝胶电泳(2-DE)的可重复性及分辨率。方法:临床手术中抽提先天性核性白内障患儿混浊晶状体组织蛋白,采用固相pH梯度等电聚焦(IPG-DALT)的方法,选用12.5%的SDS-聚丙烯酰胺凝胶电泳,分别以270μl、300μl上样量对晶状体组织总蛋白(crystallin)进行分离,并用银染显示分离效果。结果:上样量为270μl、12.5%的分离胶对晶状体总蛋白的分离效果较好,上样量为300μl、12.5%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93.17%。结论:采取适当的制备方法及合适的上样量可以充分分离人混浊晶状体蛋白质,并可以得到较好的分辨效果,但重复性还有待于进一步提高。
Objective: To study the repeatability and resolution of congenital nuclear cataract cloudy lens protein two-dimensional electrophoresis (2-DE). Methods: Lens proteins form 6 years old congenital nuclear cataract child were separated using 2 DE using series SDS-PAGE density (12.5 %) and silver staining methods. Results: Crystallin could be well separated from 270μl of total protein sample using 12.5% SDS- PAGE. High molecular weight proteins could be well separated from 300 μl sample using 12.5% SDS- PAGE. The matching rate was 93.17%. Conclusion: Using IPG-DALT method, electrophoresis provide better analysis for the lens protein separation, but the data repeatability should be further improved.
出处
《新疆医科大学学报》
CAS
2008年第1期34-35,38,共3页
Journal of Xinjiang Medical University
基金
国家自然科学基金资助项目(30471864)
关键词
晶状体
蛋白质
双向凝胶电泳
lens
protein
two-dimensional electrophoresis