our degas machine was broken today, and it will be repaired at next month. but recently i will use FPLC to purify protein, so how can i degas the buffer, milli-Q, and 20% EtOH?
Hi there,
Sonication bath or helium bubbling system are alternatives.
If you use a vacuum filtration flask to filter your buffers, water and ethanol at room temperature, then put them in the refrigerator and also perform the chromatography in the refrigerator, the degassing that occurs during filtration will be sufficient for the FPLC.
thank you, Dominique and Adam. we do not have Sonication bath or helium bubbling system, so Adam's idea maybe suitable for me. because previously i found after the buffer was filtered, i put it in 4 degree overnight, and i continue to degas at next day, but there is no air bubble during degassing, and in the cap of the bottle, there were much water vapor, this may means buffer finished degassing in cold temperature. but i do not know why.
Hi Zixun, gases have higher solublity in cold water - so you will not see the same degassing effect in your buffer after cooling it down to 4 degrees. You can leave the buffer at the vacuum system after filtration to degas and I also add a magnetic stirrer at medium speed during vacuum degassing to speed up the process. Good luck!
- Badges
- Science method