How to store SDS-PAGE for fluorescent imaging without fixatives?
I am running protein samples on SDS-PAGE that are tagged with fluorophores. If I place the gel in a destain solution of 60%water/30%methanol/10% acetic acid, the chemistry reverses and I lose the fluorophore. Is there an alternative storage solution I can use to get my gel to the imager to capture the fluorescence without chemical fixatives? After imaging, I then proceed to Coomassie Blue staining in acetic acid and methanol, at which point I am not worried about losing the fluorophore.
Adam B Shapiro
If you image the gel immediately after it finishes running, it should not be necessary to use a fixing agent. The bands will not diffuse significantly in a few minutes.
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