Optimal parameters for laccase-mediated destaining of Coomassie Brilliant Blue R-250-stained polyacrylamide gels

Detergent-mediated destaining of coomassie brilliant blue-stained SDS polyacrylamide gels.

A simple and one-step detergent-mediated destaining procedure for SDS Polyacrylamide gels for proteins is described. Suspension (5%, w/v) of a commercially available household detergent, Vim Ultra, has been found to be very efficient in destaining polyacrylamide gels without interfering with the resolution of proteins. As compared to the routinely used solvent (methanol-acetic acid-water)-media...

Rapid Coomassie blue staining and destaining of polyacrylamide gels.

Repeated probing of western blots obtained from coomassie brilliant blue-stained or unstained polyacrylamide gels.

For several reasons, it can be desirable to probe a filter obtained after Western blotting of polyacrylamide gels with different antibodies. On the one hand, protein amounts may be limiting and re-probing of a protein filter may be the only way to identify different antigens. On the other hand, in the case of two-dimensional (2-D) gel electrophoresis, exact reproducibility of the protein patter...

Removal of coomassie blue precipitates from polyacrylamide gels.

During staining of polyacrylamide gels, varying amounts of Coomassie blue may precipitate on the surface of the gels. This background obscures faint bands and is esthetically unappealing. The problem is worse when gels are stained for longer periods of time or when the stain has not been recently filtered. Rinsing a gel with methanol efficiently and quickly removes this background. Basically, a...

Criticism of the use of Coomassie Brilliant Blue G-250 for the quantitative determination of proteins.

The quantitative determination of proteins in biological fluids, using Coomassie Brilliant Blue G-250, was evaluated. Compared with the biuret method, the Coomassie Blue G-250 method needs a much shorter time for analysis and has a greater sensitivity. The sensitivity of the dye for albumin is significantly greater than for globulins. The standard curves for the biuret method are more linear th...