Hello,
As a part of my ongoing experiment, I run Gelatin-SDS-PAGE zymography for semi quantative coparitive study of MMP 9 and 2 on my samples.
The samples are unconditioned (not treated with PMA: phorbyl myristate acetate) cell culture supernatant which include 5% FBS.
The problem I'm encountering is that eventhough my standard marker is giving me a bright sharp band, my samples are giving me smears.
Since more or less this is a specific technique for gelatinases, and I am aware of the presence of Latent (proenzme) gelatinase in the sample, I would thank if anyone could advise me what to do to emit the smear resulting sharper bands.

Hi, You could try to run the samples at a lower voltage than usually recommended. And it also depends on how viscous your samples are. Another option is to de-stain the Coomassie more slowly. Hope it helps!