Detection of radioactively labeled proteins is quenched by silver staining methods: Quenching is minimal for 14C and partially reversible for 3H with a photochemical stain

Silver staining methods for protein detection in polyacrylamide gels have a quenching effect on autoradiography and fluorography. This effect was quantitated for proteins in two-dimensional gels by microdensitometry using a computer equipped with an image processor and by scintillation counting of proteins solubilized from the gels. The original histologically derived silver stain had a quenching effect that was severe and irreversible for ³H detection and moderate for ¹⁴C detection. A silver stain based on photochemical methods had minimal quenching of ¹⁴C detection and less of a quenching effect than the histological stain for ³H detection. The ³H quenching effect was partially reversible for the photochemical stain.