Bcl 3 polyclonal antibody

Testosterone propionate, estradiol benzoate and olive oil were purchased from Aladdin Co., Ltd. (Shanghai, China). NAF (>99%) and HJZ-12 (>95%) were synthesized in according with the previous methods (Huang et al., 2015 (link)). Pierce BCA Protein Assay Kit was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Anti-α-smooth muscle actin (α-SMA, 1/400) was purchased from Sigma-Aldrich (MO, United States). Anti-β-actin antibody (1/5000), Bcl-3 polyclonal antibody (1/500) and RRS1 polyclonal antibody (1/500) were purchased from Bioworld Technology (Inc., United States). Rabbit monoclonal anti-proliferating cell nuclear antigen (PCNA, 1/1000), and anti-FGFR3 antibody (1/1000) were purchased from Abcam (Cambridge, MA, United States). Anti-Bmi-1 antibody (1/1000) and anti-cleaved caspase 3 (1/1000) were purchased from Cell Signaling Technology (Beverly, MA, United States). All reagents were prepared fresh before use.

The total protein of concentration of each tissue or cell sample was determined by the Pierce BCA Protein Assay Kit (Waltham, MA, United States). 12% SDS-polyacrylamide gel and polyvinylidene-difluoride (PVDF) membranes (Millipore, Burlington, MA, United States) was used, and the membrane was blocked for 1 h at room temperature with 5% non-fat milk. The primary antibodies was incubated overnight at 4°C, including rabbit monoclonal anti-PCNA, anti-FGFR3 antibody; anti-β-actin antibody (1/5000), Bcl-3 polyclonal antibody and RRS1 polyclonal antibody (1/500; Bioworld Technology, Inc., United States); and anti-Bmi-1 antibody and anti-cleaved caspase3 (1/1000; CST, Beverly, MA). Then, the HRP-conjugated anti-mouse or anti-rabbit IgG (1/5000; Sigma–Aldrich MO, United States) was incubated for 1 h at room temperature. The enhanced chemiluminescence (ECL) western blotting detection reagents (GE Healthcare Biosciences, Pittsburgh, PA, United States) was used to get the band. The band was analyzed by ImageJ software.