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[jmafs]

Hi All,

I was wondering if anyone had thoughts on how to increase signal for MMPs in zymography.  I understand that one can either add more protein, or incubate the enzymes longer before staining, but are there any other factors that could lead to a reduction in signal?

Normally, I have read that researchers use 1-10 ng of MMP-2 standard in gelatin zymograms and get good bands, however I have found in my application that I need to increase this number to about 30ng.  This is after an overnight incubation as well, recommended for higher sensitivity.

This has been my go-to protocol.  

Any thoughts are much appreciated!

[protolder]

Hola, only one thing and excuse me if you done it, I remember that  the sample buffer for zymograms is withouth reducer and the samples are loading without boiling as the protocol says. Good luck