So recently for a few experiments I had to do zymography. I would like to check a few MMP biomarkers with respect to the experiment. I did it a couple of times and I have results, but they're not that clear. I have a few smears, but I need the degradation bands, which I can visualise properly.
as well as what recommended concentration would you people use for the sample?
as well as what tools I can use to analyse the results for the same as in how I quantify my results.
It does not appear that there was successful clearance of the gelatin by sample enzyme. The first step is getting good gelatin dispersal when casting the gel. You need a high bloom number gelatin. At 8% polyacrylamide, your gelatin appears to have electrophoresed down below the sample. The dark protein bands should not be visible. I use 10 %, some people use 12%. This is the first step to change.
see Materials and methods:
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