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Coomassie Brilliant Blue (CBB) staining

Posted on Thursday, March 22, 2007

Description
Coomassie staining is used for Acrylamide gels (1D or 2D) prior to mass spectrometry

Procedure
Prior to Coomassie Blue staining, proteins are usually
-fixed for >30 min in 20% TCA.
-After a brief (5 min) washing step with 40% ethanol and 10% acetic acid,
the gel is saturated with the dye solution (0.1% BBB R-250 in 40% ethanol and 10% acetic acid) for 3 hours, and destained (however not completely!) two times 20 min each with 40% ethanol and 10% acetic acid until the background is no longer stained dark blue. Then the gel is immersed in 500 ml of distilled water containing a few ml of acetic acid for up to 48 hours to decrease background staining and to increase sensitivity. Detection limit of this stain is better than one ug protein/spot.


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Submitted by: wild1

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