|SYPRO Orange stain (Molecular Probes, Inc.) is a novel fluorescent dye for the detection of proteins separated by SDS or native polyacrylamide gel electrophoresis. The staining procedure is simple, rapid and sensitive. (An excellent report describing in detail the use of SYPRO Orange to stain protein gels can be found in Malone et al., Electrophoresis, 2001, 22, 919-932.)
The detection limit for SYPRO Orange-stained proteins using a DR transilluminator is around 2 - 3 ng both by eye and using Polaroid 667 film. This level of sensitivity, especially by eye, is significantly greater than that obtained using a UV device (about 20 ng).
|Molecular weight standards were subjected to SDS PAGE. After electrophoresis, the gel was stained with SYPRO Orange and photographed on a Dark Reader transilluminator using Polaroid 64 color film.
The protein load on this particular gel ranged from 120 ng to 15 ng of protein per band (left to right). The inset shows the same gel stained with Coomassie Blue.
Not only is the sensitivity of SYPRO Orange much greater than that of Coomassie Blue, but the entire procedure is complete in 20 minutes. After Coomassie staining, on the other hand, the gel has to be de-stained for several hours.
|Link to SYPRO Orange at Molecular Probes|
Clare Chemical Research