I need very rapidly to see my protein on the gel. How to stain it with Coomasie, and destain in the shortiest time possible?

Normally I do coloration for O/N, and decoloration with acetic acid solution during 4 hours, in changing solution every 1 hour .
How to do all this just in several hours?

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Alesia...ur best bet is ponceau red. It stains in 15 mins and destains quickly too... Coomassie is more permanent. Ponceau red...Sigma has a ready made solution...all that u have to do is mix it in water.
good luck

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we usually stain the gel with a 10% acetic acid solution containing 0,06 g/l coomassie G250 for 30 minutes. The proteins appear during the staining!. We destain with 10% acetic acid solution for 1 hour.

Good luck!

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hi alesia,

one technican in my old lab used a microwave for rapid staining and destaining. she cooked the gel in the staining solution for 5-10 min and destained the gel by cooking with destaining solution twice for 5-10 min. but you should be careful and just using microwave suitable boxes with a lid. and consider please that cooked acetic acid smells not so funny....
and another technican told me, that she used just aqua dest for destaining in the microwave...
but i must add that i have never accomplish this both methods by myself. i stain o/n and so on..

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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.

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You can use the very quick method: microwave for 1 min at max with stain sol (Coomassie) and destain same way, BUT, you might get a not so nice gel. If is just to see if you have something in there, it works, if you need a really nice pic, it doesn't.

Anyway, I'm using now a solution that contains Coomassie G250 instead of the regular R250. Stain for 15 mins, shaking at RT, and destain for about 1 hour, changing destainer every 15 mins or so. If you leave in destainer overnight, you'll get completly colorless background.

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you can buy Coomassie stain from Bio-Rad (i think) that are much faster staining and destaining them the lab made coomassie

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Yep....

We destain the gel afterwards just in water in a microwave (on high) for one minute (depends on your volume) or till it starts boiling. Remove the now blue stained water, add fresh water and repeat the microwave procedure till you think you`ve destained enough (always remove stained water.. )..

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the bio-rad biosafe coomassie is ok, but from experience the pierce imperial protein stain is anytime better. both are fast but the pierce seems to deal with low abundance proteins much better than bio-rad which does not seem to stain them well, especially in a 2D gel.

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This is a very quick and easy Coloidal-coomasi stain method, you usually don't need to destain at all, just wash quickly in water. You can also skip out the TCA fixation step if you don't need to quantify and are just looking quickly for bands.

Coloidal Coomasi G250
Method of Neuhoff et al. Uses MEOH which is toxic.

• Fix gel in 50ml of 12% Tricholroacetic acid TCA (store at 4°C) for 1 hour.
• Prepare staining solution. Make up 80ml of 0.1% w/v CBBG-250, 2% W/Phosphoric acid, 10% Ammonium Sulphate, 20ml MEOH. Adjust to a final volume of 100ml with dH20.
• Add 30ml of staining solution for between 2-12 hours.
• Wash briefly in 25% MEOH (optional, will destain bands slightly)
• Destain in dH20 for between 0.5-3 hours, adding tissue and changing the water.

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