Silver Staining of Sulfated Glycosaminoglycans: A Staining Method for Visualization of Sulfated Glycosaminoglycans in Polyacrylamide Gels
Transkript
Once the run is complete, disassemble the cassette, and carefully extract the gel into a large, clean container filled with deionized, filtered water. Then, discard the water and stain the gel in Alcian blue staining solution for five minutes. Discard the Alcian blue stain, and quickly wash two to three times with deionized, filtered water until most of the Alcian blue staining solution has been removed.
Stain the gel in silver nitrate staining solution in a fresh, clean container for 30 minutes. Then wash two to three times for 30 minutes each in deionized filtered water, to fully remove the silver staining solution. Discard the water, and add developing solution.
Carefully observe the gel for the appearance of bands. Depending on the quality of the stain and the mass of the sample loaded, development can take anywhere from a few seconds to several minutes. As soon as the desired bands are visible, immediately discard the developing solution, and wash the gel briefly with stop solution.
