Goat polyclonal anti rabbit hrp

A total of 2 × 10⁵ cells were lysed in SDS loading buffer, heat denatured, and proteins were separated by electrophoresis using 12% SDS resolving gels. The proteins were transferred to nitrocellulose and probed for BMI‐1, p16, and β actin expression using mouse anti‐p16 (Millipore MAB4133; Watford, UK) at 1 in 1000 dilution (1 μg/mL), mouse anti‐BMI‐1 (Millipore Clone F6 Cat No 05‐637) primary antibody at 1 in 1000 dilution, and rabbit polyclonal anti‐β actin (Abcam ab8227, lot 712923, 0.65 mg/mL) primary antibody at 1 in 5000 dilution. Secondary antibodies were used at 1 in 10000 dilutions and consisted of goat anti‐mouse HRP (Jackson Immuno 115‐035‐062) and goat polyclonal anti‐rabbit HRP (Sigma A0545). ECL reagent was used to visualize proteins as directed by the manufacturer (GE Healthcare RPN2209; GE Healthcare, Amersham, UK).

The primary antibodies and reagents used for Western blot and immunoprecipitations include rabbit polyclonal anti-HBc antibodies (Dako), mouse monoclonal anti-HBc (clone Hyb3120, CosmoBio), anti-β-actin (Abcam), rabbit monoclonal anti-PRMT5 (Abcam), anti-MEP50/WDR77 (Abcam), anti-histone H3 (Abcam), anti-alpha-tubulin (Abcam), anti-phosphoserine (Abcam), anti-symmetric dimethyl arginine (SDM-R) (Cell Signaling), anti-monomethyl arginine (MM-R) (Cell Signaling), anti-HA (Santa Cruz), anti-Ubiquitin (Cell Signaling), anti-SmB/B’ (Sigma), mouse anti-Flag (Sigma-Aldrich), anti-HA magnetic beads (ThermoFisher), and anti-Flag magnetic beads (Sigma). Secondary antibodies include goat polyclonal anti-mouse horseradish peroxidase (HRP) (Sigma-Aldrich) and goat polyclonal anti-rabbit-HRP (Sigma-Aldrich). Antibodies for immunofluorescence analysis (IFA) include mouse monoclonal anti-HA antibody conjugated with FITC (Sigma-Aldrich).