wr1015 Posted April 11, 2007 Posted April 11, 2007 is it ok to store SDS-PAGE gels in ddIH20 overnight and possibly longer after staining?
ecoli Posted April 11, 2007 Posted April 11, 2007 If you're talking about Coomasie stain, you can leave it coomasie overnight, and destain + dry the next day. You can save gels before running them. Wrap them in kim wipes, water and finally seran wrap or plastic. they can stay at 4 deg. C for up to a month.
ic3reyes Posted April 11, 2007 Posted April 11, 2007 Standard protocol in our lab is to put them in ddH2O for awhile after destaining to let them rehydrate. Doing this before fixing in gel drying film seems to reduce the likelihood of the gel tearing as it dries. I have sometimes left mine in water for up to a week without any damage.
wr1015 Posted April 11, 2007 Author Posted April 11, 2007 thanks for the replies, looks like i should be ok then
ecoli Posted April 11, 2007 Posted April 11, 2007 Standard protocol in our lab is to put them in ddH2O for awhile after destaining to let them rehydrate. Doing this before fixing in gel drying film seems to reduce the likelihood of the gel tearing as it dries. I have sometimes left mine in water for up to a week without any damage. Just out of curiosity, why do this when you can just dry them right away. It doesn't really take THAT long, and once it's dry, it last for years (AFAIK).
ic3reyes Posted April 12, 2007 Posted April 12, 2007 Well, reason #1 is that my PI is a "dogma fanatic", and that's the way they did it when he was a postdoc, so that's the way we do it in our lab . I also think we've had issues with massive gel cracking, maybe due to the low humidity since we're in a desert. But you're right, you don't really need to rehydrate SDS-PAGE gels after destaining before you dry them.
CharonY Posted April 16, 2007 Posted April 16, 2007 Actuallly waterin the gels has some merits. There are number of coomassie staining protocols and in most of them siginifcant shrinking of the gel occurs during fixation and staining. Those gels easily crack if you dry it, especially if the ambient air is also dry (in deserts, of course, but most labs have a dry air due to their ventilation systems). Regarding storage, if you use water I fond that I still could use mass-spec on most gels even after half an year or longer, though mould could be problem till then. In fact, you can also restain your gel several times, if need be.
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