These days I found a strange thing happened during SDS-PAGE gel running. I found the blue dye is diffusing, forming big smear. At the same time, the pre-stained protein marker was sperated well in the resolving gel, because I can clearly see the ladders. After SDS-PAGE gel running, when I finished western blot, I can see my protein of interest as a single band, with expected size. So it seem that this blue dye diffusion does not affect SDS-PAGE.
But this phenomenon only appears recently, before this, the blue dye forms a very nice front line during gel running, it is very thin, crossing the whole gel.
I have changed everything such as loading buffer, running buffer, also, thoes reagent that used in preparing gel. But I still observed the same thing.
Did anyone experience this before? Although it doesn't affect the result, I'm very curious to know the reason.
Thank you very much:)
smear of loading buffer observed
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