Hello. I have some problem in SDS-PAGE.
My sample condition was this.
1. Ammonium sulfate gradient(20~40%)
2. Dialysis(1kDa)
3. After filter then freezing dry.
4. Dissolve using phosphate buffer
5. SDS-PAGE running.
After SDS-PAGE done. Before staining , I can see white and milk color band nearby 4kda. And half gel did staining and another half gel did zymography.
In zymography, white and milk color was lysis my indicator strain. But in staining, white and milk color band didnt stain. Is this not protein? Ah my bacteria is bacillus subtitles.
Hi Woobin, if you see any bands on SDS PAGE before staining with Coomassie then you have a problem.
How does the bromophenol blue dye front run? Is it sharp or is it diffuse? If it is diffuse you are loading too much protein or your sample has too much salt in it.
Than you answer to me. When I run sds page that time dye shape loos like Ellipse. And after more running sds page, dye shape change little sharp shape.
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