Increasing IP Yield

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DatUVa
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Increasing IP Yield

Hello Folks,

I am attempting to IP a protein from cell culture in sufficient quantity to see it by silver staining. Although I can successfully IP enough to see by Western, the yield is still too low to see by silver stain. I expect I'll have to use several plates in order to get enough protein. My question for people who have done this is, how do you go about pooling the plates? Do you add more protein to the IP, and scale up the other reagents accordingly? (beads, antibody) Do you do several individual IPs and somehow pool them after eluting the protein? The problem with that is that there is a maximum volume that can be loaded into a well of a gel.

Thanks for any suggestions.