Hi All
We ran out of Bromophenol blue in the lab. Is it plausible to use trypan blue instead? I plan on using the proteins separated on the SDS-PAGE gel downstream in a Western blot. Will it affect the transfer?
Trypan blue will not affect your proteins. I don't know whether it works well to obtain a sharp front in SDS-PAGE. But judged from its chemical structure with 4 sulfonic acid groups instead of one in bromophenol blue it should work.
Peter
Not sure about the Trypan Blue, but you don't have to have dye in your sample for the gel to run well, or for Western blot; it is just convenient for loading and knowing when the gel is done. If you run the gels routinely with the same time, voltage, etc., stop the run when you normally do. Or you can follow the progress of the dye front in the MW ladder lane, or better yet, use prestained standards to know when electrophoresis is finished.
Hi there,
It might stain proteins in some way...
Article Trypan blue staining of proteins fractionated by acrylamide ...
The transfer of the protein from SDS-PAGE to Western blotting could be affected by Trypan blue which could complex with the protein due to its excess sulphonic acid group over bromophenol blue.
However, protein staining (with Bromophenol blue) is not very necessary while running SDS-PAGE if you have been working with the protein, you can predict the time for the full electrophoresis.
It might not affect the protein, but it might well affect your health. Investigate about the safety measures of trypane blue...
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