摘要
目的:探讨肺腺癌组织中HSP70多肽复合物分离及纯化的可行性方案。方法:采用裂解液裂解、超声破碎、ConA-sepharose亲和层析和DEAE阴离子交换层析方法,从肺腺癌组织中分离纯化HSP70蛋白复合物;通过SDS-PAGE电泳、Westernblot检测获得蛋白;运用Bradford法对其进行定量分析。结果:通过该实验方法可以从1g肺癌组织中获取110μg左右的HSP70多肽复合物,其相对分子质量为70000。结论:采用ConA-sepharose亲和层析和DEAE阴离子亲和层析相结合的蛋白纯化法可以获取高纯度的HSP70多肽复合物。
Aim:To explore a method of separating and purifying HSP70 peptide complex from human lung adenocarcinoma.Methods:HSP70 peptide complex was separated and purified from human lung adenocarcinoma tissue by means of cell lysis,ultrasonication,ConA-sepharose column,and DEAE ion-exchange chromatogaraphy.The purified fractions were tested by SDS-PAGE and Western blot.And quantitative analysis was made by Bradford.Results:About 110 μg HSP70 peptide complex was obtained from 1 g lung adenocarcinoma tissue by this experimental method,with relative molecular weight of 70 000.Conclusion:The high purity HSP70 peptide complex could be obtained by ConA-sepharose column and DEAE ion-exchange chromatogaraphy.
出处
《郑州大学学报(医学版)》
CAS
北大核心
2012年第6期753-755,共3页
Journal of Zhengzhou University(Medical Sciences)
基金
国家自然科学基金资助项目30600490
河南省教育厅科技创新人才支持计划基金资助项目2008HASTIT028
河南省教育厅自然科学研究基金资助项目2008A320018
关键词
肺腺癌
HSP70多肽复合物
蛋白分离纯化
lung adenocarcinoma
HSP70 peptide complex
protein separation and purification