After hydrolysis of type II collagen using proteolytic enzymes, I got different molecular weight bands in SDS-PAGE. I used molecular weight cutoff membrane (100,50,30 kDa) to separate. This method seems like improper and also i got 70,55, 25, 15 kDa which cant separate by this MWCO membrane. I also used column chromatography (Sephadex G-25), it is not working properly.
Bruce Pratt
If you don't need too much material, just cut the bands out of an unstained gel and elute out the protein.
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