Peroxidase conjugated anti mouse secondary antibody a 4416

All antibodies were used at a 1:1000 dilution in 5% non-fat milk for western blot. Anti-ERG antibody [EPR3864 (2)] and (C-20) were purchased from Abcam and Santa Cruz, respectively. Anti-SPOP antibody (16750-1-AP), Anti-BRD2 (22236-1-AP) and anti-BRD3 (11859-1-AP) antibody were purchased from Proteintech. Anti-BRD4 antibody was purchased from Cell Signaling (13440) and Bethyl laboratories (A301-985A-M). Anti-Cullin 3 (2759), anti-GST (2625), anti-DEK (13962s), anti-SRC3 (2126), anti-Cleaved Caspase-3 (9664), anti-Cleaved PARP (5625), polyclonal anti-Myc-Tag antibody (2278) and monoclonal anti-Myc-Tag (2276) antibodies were purchased from Cell Signaling. Polyclonal anti-Flag antibody (F-2425), monoclonal anti-Flag antibody (F-3165, clone M2), anti-Tubulin antibody (T-5168), anti-Vinculin antibody (V-4505), anti-Flag agarose beads (A-2220), anti-HA agarose beads (A-2095), peroxidase-conjugated anti-mouse secondary antibody (A-4416) and peroxidase-conjugated anti-rabbit secondary antibody (A-4914) were purchased from Sigma. Monoclonal anti-HA antibody (MMS-101P) was purchased from Biolegend. Anti-GFP (8371-2) antibody was purchased from Clontech. Anti-Cullin 1 (SC-17775), anti-AR (N-20), anti-TRIM24/TIF1α (C-4), polyclonal anti-HA (SC-805) and anti-p27 (SC-528) were purchased from Santa Cruz. Anti-Xpress (46-0528) antibody was purchased from Invitrogen.

Cells were lysed in EBC buffer supplemented with protease inhibitors cocktail and phosphatase inhibitors, and the protein concentrations were measured as described above. The lysates (40 μg protein) were then resolved by 10% SDS-PAGE at 130 V for 80 min, immunoblotted with indicated antibodies at 4 °C overnight, washed 4 times with Tris-buffered saline with 0.1% Tween-20 (TBST), incubated with secondary antibody for 1 h at room temperature, and then washed 4 times with TBST buffer. Anti-p65 (#8742, 1:1000) and E2F1 (#3742, 1:500) antibodies were purchased from Cell Signaling Technologies. Anti-HA antibody (#901513, 1:1000) was purchased from Biolegend. Anti-Flag (F3165, 1: 5000), antivinculin antibody (V-4505, 1: 50000), peroxidase-conjugated antimouse secondary antibody (A-4416, 1:3000), and peroxidase-conjugated antirabbit secondary antibody (A-4914, 1:3000) were purchased from Sigma. All primary antibodies were diluted in 5% bovine serum albumin (BSA) in TBST buffer, and secondary antibodies were diluted in 5% nonfat milk in TBST buffer.