GFP-tagged proteins were recognized with a monoclonal α-GFP antibody (Roche; 1:1000) and arginine methylation was recognized with a monoclonal α-mono and dimethyl arginine antibody (Ab412, Abcam; 1:500), followed by a secondary α-mouse-horseradish peroxidase (HRP) antibody (GE Lifesciences; 1:5000). Slr1-TAP was recognized with an HRP-conjugated goat anti-Protein A (PrA) antiserum (Rockland; 1:10,000). Ribosomal protein Rps3 was recognized with a polyclonal rabbit antibody against Rps3 (1:1000). Proteins were visualized through enhanced chemiluminescence (Pierce) and autoradiography.
Anti-rabbit IgG (H+L)-HRPO and anti-mouse IgG (H+L) HRPO (Dianova) secondary antibodies were used for sucrose density gradient fraction immunoblots. The signals were detected with Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and the FUSION-SL chemiluminescence detection system (Peqlab) and the Western blot analyses were quantified using the Image StudioLite software (LI-COR).
Anti-rabbit IgG (H+L)-HRPO and anti-mouse IgG (H+L) HRPO (Dianova) secondary antibodies were used for sucrose density gradient fraction immunoblots. The signals were detected with Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and the FUSION-SL chemiluminescence detection system (Peqlab) and the Western blot analyses were quantified using the Image StudioLite software (LI-COR).