Hi,
I want to add about 5 CMC DDM detergent in my Size exclusion buffer. Is it advisable to add detergent and then degass by vacuum or degass the buffer first and then add DDM dropwise slowly??
Hi there,
A filtration under vacuum is enough to degass sufficiently for FPLC. Break the vacuum right after filtration.
If you want to degass by vacuum, you'll have to do it before you add detergent, otherwise the solution will foam. Then add the detergent from a concentrated stock and mix gently, so as not to dissolve air.
Alternatively, degass in a sonicating water bath, this can be done with detergent.
The addition of detergent should be done after degassing the buffer. Else, the whole buffer will foam while degassing with a vacuum pump. However, if the detergent is added after the degassing process, the solution should be stirred gently to allow proper mixing.
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