Does anyone have a general protocol on how to prepare cell culture media for an ELISA assay, specifically for secreted proteins in vitro?

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No title

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Ali Zohaib

Dear

We always collect the supernatant and store at -80C

Regards

Karan Wadhwa

I always aliquot and freeze culture media at -80, avoiding freeze thaw as this affects many proteins.

Ramsey Najm

Thanks so much for the help! Just one more question...

Is there a certain protocol for collecting the media? For example, it would make sense to me to add new media and allow the same amount of time in each experiment between adding new media and collecting it in order to try and have relatively controlled protein levels. Is this important or is it enough to feed 24 hours before hand, or something along those lines, and collect the media then.