The fusion protein I´m working with stains well with coomassie, but not with silver. After the fusion hydrolysis, the digestion subproducts are stained both with silver and coomassie, but I can´t detect the untreated fusion with silver.
Are all the silver staining steps being followed thoroughly and properly? What about all the reagents that you need for silver staining? I hope they're in good condition. Is it working in case of other samples too?
Hi Amanda, it's possible that your protein is binding the silver ions and preventing the reaction. This binding activity maybe it's primary structure dependent, so digestion products do stain.
Thank you all for your answers. The silver staining reagents are ok, I repeated the procedure twice with the same results. The staining works for other proteins with lower amounts in the same gel. I'm using SDS-Trcine-PAGE, so all the proteins are under denaturing conditions.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining