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I usually heat to boil in the microwave, and stain for 12 minutes. How often can I reuse this buffer?

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Since there's a lot of methanol in Coomassie stain, a significant amount would probably evaporate off each time you microwave it. Therefore, it'd be probably a good idea not to reuse it because (1) you're losing methanol, and (2) the concentration of everything else is thrown off.

Why do you boil the stain? I just pour mine at RT right on the gel and it works fine.

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Doesn't heating it up make it stain faster? Also, labs I've worked in have done both: reuse and not. My impression is that you can reuse some of it, but it's best if mixed 50/50 with fresh stain. But of course, this is all lab folklore. I'd be curious to know if there has ever been a systematic study of whether/how much re-use affects results. – Thomas Ingalls Feb 18 '12 at 21:00
I've never boiled it and it always stained my gels fine in 10-15 minutes. (Plus it takes a few minutes to boil anyways... and sniffing those methanol vapours isn't exactly good for you...) – jp89 Feb 18 '12 at 23:42
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And after I've stained the gel, I just pour it back into the container. I've used the same bottle of stain for 2+ years. Works like a charm. – jp89 Feb 19 '12 at 2:54

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