How to make serum for tissue culture?

[mark]

John HI, 
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?