Goat anti mouse horseradish peroxidase secondary antibody

Harvested proteins were first separated by 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes (Bio-Rad Laboratories, USA). The membranes were blocked with 5% nonfat milk and incubated with antibodies. The membranes were subsequently incubated with a goat anti-mouse horseradish peroxidase secondary antibody (Sigma, USA). The protein complex was detected using enhanced chemiluminescence reagents (Pierce, France). Endogenous beta-actin was used as the internal control. LCN2 antibody was purchased from R&D Systems, USA. AR, PSA, SLC45A3, FKBP5 and NKX3.1 were purchased from Santa Cruze, USA.

Cellular proteins were extracted and separated in SDS-PAGE gels, first separated by 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes (Bio-Rad Laboratories, USA). The membranes were blocked with 5% nonfat milk and incubated with a mouse anti-AMFR polyclonal antibody at a dilution of 1:500 (Abcam, USA), a rabbit anti-activated NOTCH1 antibody (Abcam, USA), or a mouse anti-beta-actin monoclonal antibody at a dilution of 1:1000 (Sigma, USA). The membranes were subsequently incubated with a goat anti-mouse horseradish peroxidase secondary antibody (Sigma, USA). The protein complex was detected using enhanced chemiluminescence reagents (Pierce, France).

Harvested proteins were first separated by 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes (Millipore, USA). The membranes were blocked with 5% nonfat milk and incubated with a mouse anti-PLD1 polyclonal antibody at a dilution of 1:500 (Cell signal, USA) or a mouse anti-beta-actin monoclonal antibody at a dilution of 1:500 (Sigma, USA). The membranes were subsequently incubated with a goat anti-mouse horseradish peroxidase secondary antibody (Sigma, USA). The protein complex was detected using enhanced chemiluminescence reagents (Pierce, France). Endogenous beta-actin was used as the internal control.