Rabbit polyclonal 14 3 3 pan antibody

Rabbit polyclonal C1orf64 antibody (Novus Biologicals, Littleton, CO, USA) and rabbit polyclonal SPDEF antibody (Life Technologies) were applied at 1:250 dilutions. Rabbit polyclonal AR antibody (Active Motif, Carlsbad, CA, USA) and rabbit polyclonal 14-3-3 (pan) antibody (Millipore, Temecula, CA, USA) were used at 1:1000 and 1:5000 dilutions, respectively. Rabbit monoclonal PIP antibody (Novus Biologicals) was applied at 1:1000 dilution. Mouse monoclonal α-tubulin antibody (Sigma-Aldrich) was applied at 1:2000 dilution to assess loading. Protein concentrations were measured using the BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA) and a total of 20 μg of each cell lysate was used for immunoblotting. Immunoblot imaging and analysis of band densities were performed by a C-DiGit Blot Scanner (LI-COR, Lincoln, NE, USA). Western blots were performed in three replicates and the average fold changes were shown.

Rabbit polyclonal SRARP (C1orf64) antibody (Novus Biologicals, Littleton, CO, USA) and rabbit monoclonal HSPB7 antibody (Abcam, Cambridge, MA, USA) were applied at 1 : 250 and 1 : 1000 dilutions, respectively. Rabbit polyclonal 14‐3‐3 (pan) antibody (Millipore, Temecula, CA, USA) was used at a 1 : 5000 dilution. Rabbit monoclonal antibodies for ERK1/2, phospho‐ERK1/2 (Thr202/Try204), Akt (pan), and phospho‐Akt (Thr308) were obtained from Cell Signaling Technology (Danvers, MA, USA) and applied at 1 : 1000 dilutions. Mouse monoclonal α‐tubulin antibody (Sigma‐Aldrich) was applied at a 1 : 2000 dilution to assess loading. Protein concentrations were measured using the BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA), and a total of 30 μg of each cell lysate was used for western blotting. Western blot imaging and analysis of band densities were performed by a C‐DiGit Blot Scanner (LI‐COR, Lincoln, NE, USA). Experiments were performed in three replicates, and mean fold changes are presented.