Cytochemical method to localize acidic nuclear proteins

Fast green FCF was used to localize acidic nuclear proteins in sections of young flower buds of Limnophyton obtusifolium (L.) Miq. After extracting nucleic acids, the slides were stained at hydrogen ion concentrations ranging from pH 2.6 to 9.0. At pH 5.0 and 8.0 staining is confined to the nucleus with no cytoplasmic reaction. Staining intensity is greater at pH 5.0 than at pH 8.0. The proteins responding to fast green at pH 8.0 are basic proteins. The positive reaction at pH 5.0 is attributed to acidic nuclear proteins. These findings are confirmed by control preparations. Acetylated slides and slides treated with 0.25 N HCl were unstained at pH 8.0 but staining at pH 5.0 was undisturbed. Dilute alkali (0.003 N NaOH) reduced staining intensity at pH 5.0 but had no effect at pH 8.0. Methylated slides did not stain at pH 5.0, but at pH 8.0 staining was unaffected. Deamination blocked staining at both pH's. It is concluded that fast green at pH 5.0 specifically binds with acidic nuclear proteins.