小中大确实见过很多人这样操作的,高盐再生或者预平衡,平衡液平衡,上样、洗杂、洗脱,高盐再生,0.5M NaOH是消毒(Clean)。
附:
All cation-exchange chromatography runs were conducted as the following
procedures: Columns were equilibrated with 2 column volumes
(CVs) of preequilibration buffer (equilibration buffer with added 1 M
NaCl) and 2 CVs of equilibration buffer in tandem; columns were then
loaded with protein solution and followed by 2 CVs of equilibration buffer
as postload wash; a linear gradient elution was performed from 0 to 100%
elution buffer in 10 CVs; after that, columns were regenerated with preequilibration
buffer and cleaned with 0.5 M NaOH plus 1 M NaCl; and columns
were stored with 0.1 M NaOH at room temperature. All runs were
performed at room temperature and the flow rate was 180cm=hr.
但是我不是太明白3个问题
1,如果消毒后保存在0.1M NaOH中或者乙醇中,下次使用前是不是要先用水冲洗后,才是高盐预平衡或平衡,水冲洗前是否还需在0.5M NaOH先冲一下;
2,使用起始的时候为什么一般都采用高盐再生或者预平衡一下,这样的目的是什么?因为后面不是平衡液还要平衡的么,是为了离子能快速平衡还是其它目的。
3,最后NaOH去除杂质能力很强,那是否还需要高盐再生一下呢。