I will be treating SHSY5Y cells with amyloid beta 42 obtained from E. Coli cultures. I observed that after treatment, the media became cloudy within 24 hours indicating a contamination. I did not pass it through a sterile filter since the protein may attach to the membrane and thus reduce the working concentration required.
Is there any other sterilisation method known that will prevent contamination of the cell culture media and also minimise the loss of protein for treatment?
Thanks!!
If it is after treatment then there is very less that one can do. Those filters are designed for protein, so ideally they shouldnt affect your concentration at that great level. If you are still concerned then what you can do is dissolve purified bsa in your media, and filter it through your .22 filter , now wash that filter by passing 1X pbs through it, again pass little bit of only media, and then filter your main solution. If protein want to stick then bsa will stick, and your protein would be in better position.
Can you add ethanol to your amyloid samples and remove it after a while? you can use a vacuum centrifuge for that. Amyloids are very stable, so maybe ethanol treatment is not affecting them in any way.
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