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目的 :应用快速蛋白液相色谱 (FRLC)系统建立分离和纯化 615系小鼠肝癌细胞系Hcaf细胞膜上热休克蛋白 (HSP70 )的分离方法。方法 :用经 4 3℃热处理 8h的Hcaf细胞制备裂解液 ,用ConA Sepharose 4B和MonoQ柱进行连续分离 ,所得蛋白经电泳及Western blot进行蛋白分子量及性质鉴定。结果 :纯化蛋白经鉴定为分子量70 0 0 0的HSP70。结论 :用此层析法经FRLP可分离纯度较高的HSP70。
Objective : To establish a rapid protein liquid chromatographic (FRLC) system for isolation and purification of heat shock protein (HSP70) from the plasma membrane of Hcaf cell line 615. Methods : Hcaf cells were treated with heat treatment at 43 °C for 8 h to prepare lysates. ConA Sepharose 4B and MonoQ column were used for continuous separation. The protein was identified by electrophoresis and Western blot. Results : The purified protein was identified as HSP70 with a molecular weight of 700 0 0 . Conclusion : High purity HSP70 can be isolated by this chromatography with FRLP.