I have to perform gelatin zymography from cell supernatant. I pellet down the cells and collect supernatant. After that how should I proceed? How do I concentrate the supernatant?
you can concentrate your supernatant by centrifugation using membrane filters of an appropriate cut-off (ex amicon filters or similar).
if you want to visualize by zymography MMP-2 and MMP-9 which molecurar weight is of 62-72 KDa (active and zymogen form of MMP-2) and 82-92 kDa (active and zymogen form of MMP-9) you can use a 10 or 30 KDa cut off filter
Amicon filters are best , If you know the size of your protein, great! Choose what it suitable otherwise just go ahead with the Amicon 10000 (10 KDa) cut-off, it should retain most proteins.
All the best
To make sure the protein content, you should measured an protein level by protein assay before running to zymogram gel. Good luck..
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