How to make serum for tissue culture?
mark.haynes at mail.tju.edu
Tue Feb 17 13:44:15 EST 1998
first of all FCS is not heat inactivated when purchased and inactivation
can be achieved at 30 minutes/56C. Its quite possible that two things
are causing the hemolysis of RBCs. one is the gauge needle which sounds
unchangable the other is the length of time in shipment. Filtering serum
is unlikely to help and is guaranteed to clog filters. A 10% solution of
serum in media clogs .22 filters. Serum-free additives are available for
purchase. I think the monkey serum is a good idea. Have you compared
sera from animals that express the epitope with those that don't to
address what the carry-over may be in your system? Can you addresss the
ammount of b2m in issolated serum by a more direct approach(immunoprecip
or western?) or is this known and would it help to know it?
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