FreeStyle™ CHO Expression Medium, 6 x 1 L - FAQs

查看更多产品信息 FreeStyle™ CHO Expression Medium - FAQs (12651022, 12651014)

2 个常见问题解答

我能将FreeStyle 293表达培养基或FreeStyle CHO表达培养基直接上样至镍柱中，以用于His标签蛋白的纯化操作吗？

在使用FreeStyle 293表达培养基时，我们推荐稍早一些收获细胞，以减少宿主细胞蛋白所造成的非特异性结合，之后对培养上清液进行0.2 或0.1 µM的膜过滤，再上样至镍柱。另一方面，FreeStyle CHO表达培养基中含有的EDTA将会对镍柱起到洗脱的作用。因此在上样之前，应对这一培养基进行透析或缓冲体系交换，也可通过向上清液加入1-3 mg/L NiSO4或NiCl来螯合EDTA。另外可使用铁盐或钴盐。

Can I load the FreeStyle 293 Expression Medium or FreeStyle CHO Expression Medium directly onto a nickel column for His-tagged protein purification?

With FreeStyle 293 Expression Medium, we would recommend harvesting the cells a little earlier to reduce any nonspecific binding by host cell proteins and then filtering the culture supernatant through a 0.2 or 0.1 µM membrane before loading onto the column. On the other hand, FreeStyle CHO Expression Medium contains EDTA that will strip the nickel column. The medium can be subjected to dialysis or buffer exchange prior to loading on the column, or the EDTA can be chelated by adding 1-3 mg/L NiSO4 or NiCl to the supernatant. Iron or cobalt salts can also be used.