I am performing casein zymographies with conditioned media of cocultures of NKs cells and another cell line. In my zymography I always see a strong band of about 80 KDa. Does anyone know what protein could it be?
Ellen A G Chernoff
If your second cell type is epithelial, the 80kDa band could be matriptase released from cell membranes. Alternatively, it could be activated MMP-9. Inhibitor experiments will identify it. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1188268/
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Osmar Rivera
Ellen A G Chernoff Actually, yes. It is a human gastric epithelial cell line.
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Ellen A G Chernoff
there are small molecule inhibitors of matriptases, but I don’t know if they’ve ever been used in zymograms. I’m more familiar with MMP biochemistry. I do know both require calcium for substrate binding, but only the MMPs coordinately bind zinc ions. I always used the zinc chelator 1,10-phenanthroline on zymograms as my MMP inhibitor.
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