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目的 :探索表皮生长因子 (EGF)的制备及纯化方法。方法 :取小鼠颌下腺约 10g ,匀浆 ,离心取上清。盐析 ,离心收集沉淀。沉淀溶解于匀浆缓冲液中 ,上CM 纤维素柱。用不同盐浓度的缓冲液阶段洗脱 ,收集得A、B、C、D四个洗脱峰。用MTT法检测各峰EGF活性 ,用活性峰透析、浓缩 ,通过SephadexG 5 0柱。收集活性峰 ,浓缩 ,用聚丙烯酰胺凝胶电泳检测 ,真空冷冻干燥 ,得成品。结果 :小鼠颌下腺经匀浆 ,盐析 ,CM 纤维素柱及SephadexG 5 0柱 ,得具EGF活性、电泳为单一条带的蛋白质 ,其分子量约 6 0 0 0。得率为 1.44mg。 结论 :用盐析及层析法制备、纯化EGF ,不失为一简易可行的方法
Objective: To explore the preparation and purification of epidermal growth factor (EGF). Methods: The mouse submandibular gland about 10g, homogenate, the supernatant was centrifuged. Salting out, collecting the precipitate by centrifugation. The pellet was dissolved in homogenization buffer and loaded onto a CM cellulose column. With different salt concentrations of the buffer phase elution, collected A, B, C, D four elution peaks. The EGF activity of each peak was detected by MTT assay. The active peak was dialyzed, concentrated and passed through Sephadex G 50 column. Active peak collection, concentration, with polyacrylamide gel electrophoresis, vacuum freeze-dried to obtain the finished product. Results: The mouse submandibular gland was homogenized, salting out, CM cellulose column and SephadexG 50 column, with EGF activity, electrophoresis as a single band of protein, the molecular weight of about 600. Yield 1.44 mg. Conclusion: It is an easy and feasible method to prepare and purify EGF by salting-out and chromatography