How do you use Coomassie?

How do you use Coomassie?

Coomassie R-250 Staining Protocol

1. Prepare the staining solution containing 0.1% Coomassie R-250 in 40% ethanol, 10% acetic acid.
2. After electrophoresis, incubate 1 or 2 gels in a staining container containing 100 ml Coomassie Blue R-250 staining solution.

How long can you leave gel in Coomassie?

Note: Gel can be stained for up to 3 hours, but after 3 hours, sensitivity will decrease. If you need to leave the gel overnight in the stain, add 2 ml of 20% NaCl (w/v) in water for every 20 ml of stain. This procedure will not affect sensitivity. Wash the mini-gel with 100 ml of water for 1-3 hours.

How do you remove Coomassie gel stain?

The Coomassie stain is removed by aspiration after staining. 4. Cover the gel with ~250mL of the destain solution and allow the gel to destain with gentle agitation. The destain solution should be changed several times, removing it at each change by aspiration.

What amino acids does Coomassie blue bind to?

In acidic conditions, Coomassie dye primarily binds basic amino acids (arginine, lysine and histidine).

How do you make colloidal Coomassie?

first dissolve aluminum sulfate in Milli-Q water. thereafter add ethanol, homogenize, and mix CBB G-250 to the solution. as recently as the solution is completely dissolved, add phosphoric acid (the incorporation of the acid to the alcoholic media lets the Coomassie molecules aggregate into their colloidal state)

What does Coomassie Blue bind to?

In acidic conditions, the dye binds to proteins primarily through basic amino acids (primarily arginine, lysine and histidine), and the number of coomassie dye ligands bound to each protein molecule is approximately proportional to the number of positive charges found on the protein.

How do you dispose of Coomassie Blue stain?

The blue-colored Kimwipes did not release the stain even when squeezed dry after incubation in various salts, water, or acid solutions for five weeks. The CBB removed thus can be simply disposed as solid waste and will not leach out from solid landfills.

Can you Restain a Coomassie gel?

Yes, you can restain coommassie gels. Just wash the gel after the destain to remove any destain remaining and put the gel back into coommassie stain. You can stain and destain with coomassie as many times as you wish.