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.
Is it necessary to desain at all?
Protein willl not diffuse very much in the absence of an electric field so can you image the wet ( or even dried) gel direct from the electrophoresis run? or is there a very long delay time before imaging
I need to take the gel to a different building for the imaging, so as long as I wet it (maybe running buffer?) it should only be about a half-hour delay before I take it back for Coomassie staining.
Dear Laura Rhoads
i'm agree with Paul Rutland
you can just store the gel in milliQ water. it is stable.
Staining and destaining with methanol/acetic acid is the old way to acheive fast results but you can stain and destain gels also with water based stains.
you can find an example of this in the following video on my blog:
https://www.blogger.com/video.g?token=AD6v5dwhmY5GkO9_ezq3Va_Tfqi6u0ys99hBWy2OxXI0esS7px1EdB64LQXElfRjWFrifwHtPdDxUERBgln233r-2MAurJXhLDWcqkP1X5Gr1gSvjYZLKNS5fdeNossEHQrrnebrxiHw
best
Manuele
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