I only have a 3,000MWCO filter column to concentrate proteins in culture media. Is there a way I can alter the protocol to still include 80kD proteins?
A 3000MWCO filter should have a pore size of 3000 Da (3kDa) and therefore capture everything larger than this. So a protein with a size of 80kDa should easily stay on the membrane. Only downside is contamination from other proteins.
To follow up on Max's response. If your media contains serum, you will end up with aa really high albumin concentration. You may want to try serum-free, or at least low serum media conditioned media if your cells will tolerate it.
Filter would work, but could be messy as indicated above and your protein could stick to the membrane. You could first try ammonium sulfate cuts to get rid of the proteins you do not want, before concentrating your protein using the filter. I really like the Amicon Ultra spin filters and Centricon Plus 70 spin filters, but depends how much media you need to concentrate (can reload the spin columns several times), contact EMDMillipore and ask for test samples for your protein.
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