Protocol for Sep-Pak C18 column (waters)
For 50-100mg column:
1.)  Condition the column.  All steps can be done quickly and the solution can be pushed through using a Pasteur Pipet bulb.
First condition with 1ml MeOH
Next 1ml 80% MeCN 20% 0.1% TFA
2ml 0.1% TFA
2.)  Acidify your sample with 0.1%-1% TFA to a pH below 3.
NOTE: Usually several hundred microliters of 0.1% TFA is needed.  For excess salt in samples more 0.1% TFA may be needed. Check using pH paper.
3.)  Load sample onto the column with a slow drip.
4.)  Wash column with 3ml 0.1% TFA.  This step should be done quickly.
pushed
NOTE: For samples containing excess salt do more washes.
5.)  Elute sample with 1ml 80% MeCN in 0.1% TFA.  Do this step with a slow drip and COLLECT the flowthrough.
6.)  Dry in speedvac.
NOTE:  For larger columns use larger volumes for washes.

版权声明：本站内容均来自互联网，仅供演示用，请勿用于商业和其他非法用途。如果侵犯了您的权益请与我们联系QQ:729038198，我们将在24小时内删除。