人肾癌组织中热休克蛋白70的纯化与鉴定分析

Purification and identification of heat shock protein 70 from human renal cancer tissue

目的探讨人肾癌组织中热休克蛋白70纯化的有效方法,并进行鉴定分析。方法应用两次亲和层析和离子交换层析获得目的蛋白,经电泳和Western-blot定性分析,应用改良的Bradford法定量分析。结果经过二次亲和层析和MonoQ柱的分离后,得到纯度较高的分子量约为70kDa的蛋白质,经免疫印迹分析证实该蛋白质即为HSP70。与其他方法相比HSP70的纯度更高,获得率接近。结论该文所述方法是一种简单有效的纯化肿瘤组织中HSP70的方法。

[Objective] To investigate the method of purification of heat shock protein 70 from human renal cancer tissue and identification of it. [Methods] HSP70 in truman renal cancer tissue were purified by a modified chromatographic method including two steps affinity chromatography and ion exchange chromatography, then qualitative analysis by electrophoresis and western blot, quantitative analysis by modified Bradford＇s method. [Results] Apparent purity proteins of Mr 70 kDa were obtained using two steps affinity chromatography followed by Mono Q column. This protein was identified as HSP70 by immuno-blot. Compared with previous method, the purity of the HSP70 is higher by our method, and ha,west yield is corresponding. [Conclusion] Our method of purifying HSP70 from cancer tissues is convenient and effective.