Rabbit anti human stat3

The protein in supernatant was extracted and quantified using bicinchoninic acid (BCA) Protein Assay Kit (Pierce Chemical Co., Rockford, Illinois, USA). Equal amounts of protein (thirty micrograms) were denatured at 98 °C for 5 min and were separated via10% SDS-PAGE gel and electro transferred to nitrocellulose membrane (Millipore, Temecula, California, USA). Next, the membranes were blocked overnight at 4 °C with 5% non-fat milk power, probed with the following primary antibodies at 37 °C for 2 h: Rabbit anti-human CTBP1(#8684), rabbit anti-human CTBP2 (#13,256), Rabbit anti-human phospho-Stat3 (#9145), rabbit anti-human Stat3(#12,640), rabbit anti-human phospho-JAK1 (#74129), rabbit anti-human JAK1 (#29,261) were purchased from Cell Signaling Technology (Boston, USA), and mouse anti-human β-actin (ab8227) were purchased from Abcam (Massachusetts, US). After washing, the membranes were further incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Cell Signaling Technology, Inc, Danvers, MA, USA) at a 1:1000 dilution for another 2 h. Finally, the protein binds were visualized using ECL kit (Biorobot, Shanghai, China) on Bio-Rad ChemiDoc MP imaging system (Bio-Rad Laboratories, Hercules, CA, USA).