How to stain /destain quickly SDS-PAGE gel? - your patents? (Sep/29/2005 )

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What does the stain solution and destain solution make up? Our stain solution contains methanol. Will the methanol make some dangers to people when heated? How do you handle with it?

Thanks!

QUOTE (HomeBrew @ Sep 30 2005, 02:48 AM)

We submerge the gel in Coomassie stain and microwave on high for 45 seconds to one minute (the amount of time you zap for is obviously volume dependant -- the idea here is to get the solution hot, not cook the hell out of it or melt the gel -- 1 min is usually about right for ~30 ml). Place the box in the hood, and let staining continue for 10 minutes or so with rocking. Decant the Coomassie stain, rinse briefly with destain solution to remove the bulk of the leftover stain, the submerge the gel in destain and microwave as before.

At this point, it is usually quite evident what the gel contains, then I routinely leave the gel in fresh destain overnight, and scan it in the morning.

I actually recall reading somewhere that you could replace the methanol in these staining and destaining solutions with ethanol to make this a less nasty protocol (the methanol is very volatile and smells), but I haven't tried it yet.

-ThreeHeis-

yes, methanol is toxic. you should not inhale it.
That's why you should not boil the gel, but rather warm the solution.
you could warm the solution in a bottle in a water-bath, and add the solution to the gel under a ventilated hood.

-Missele-

We have a staining solution made with picric acid and coomassie brilliant blue R250.

You can easly see the protein during the coloration (10min), and you can also use hot water for cleanning the background (30min)....

-Marica-

but why would you want to use picric acid (with the storage, handling and disposal considerations) if you don't have to.

-mdfenko-

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