Is it possible to double stain my gel with silver stain and a coomassie dye stain?
Yes. Our researchers have stained with coomassie dye stains, both before and after staining with the Color Silver Stain. (Counter-intuitively, silver stains detect heavily loaded proteins poorly, and double-staining with coomassie dye remedies this problem). When double staining, avoid dark background by halving the silver staining incubation time or by diluting the silver solution. Consider using GelCode Blue Stain Reagent (Product # 24590 or 24592).
What can I do about high background?
Some researchers obtain lower background by omitting the stabilizer step. Omitting this step, however, will decrease sensitivity and the colors will not be as distinctive as they would be otherwise. The product instructions contains a destaining protocol that will decrease background. Because thinner (0.75 mm) gels destain more quickly than thicker gels, consider diluting the destaining solution 1:1 with water to slow down the reaction when using thin gels.
What causes horizontal streaks in a gel?
Bands that show up between 50-68 kD may be caused by beta-mercaptoethanol (BME). To avoid these bands, filter the BME or switch to dithiothreitol (DTT).
What causes vertical streaks in a gel?
Old running buffer may cause vertical streaks. To avoid these streaks, always use fresh buffer.
What incubation time should be used with a 1.0 mm thick gel?
We recommend staining the gel for 30 minutes. If the background is too dark, decrease the staining time to 20 minutes.
What is the sensitivity of GelCode Color Silver Stain?
Although detection will vary from protein to protein, this product will detect down to 0.1 ng of most proteins. The thicker the gel, the greater the sensitivity will be.
What is the shelf life of GelCode Color Silver Stain Kit?
The GelCode Color Silver Stain Kit is stable at room temperature (RT) for one year. All working solutions, except for the stabilizer, must be made immediately before each use. The stabilizer can be prepared in advance and stored at RT for several months.