Tgf β antibody

All the chemical reagents were purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China) and used without further purification or modification unless otherwise specified. Fetal bovine serum (FBS), RPMI 1640 medium, DMEM medium, and trypsin–EDTA were purchased from Gibco-Brl (Grand Island, NY, USA). Rabbit anti-mouse or anti-human PD-L1 antibody, E-cadherin antibody, Vimentin antibody, goat anti-rabbit IgG HRP and β-actin antibody were purchased from Affinity Biosciences Inc (USA), AMPK antibody, pAMPK antibody, and TGF-β antibody were purchased from Cell Signaling Technology (USA). All biochemical reagents were used without further purification.

Renal cortices were homogenized in lysis buffer (Kaiji, Shanghai, China) on ice for 30 minutes. Western blotting was performed as previously described [13 ]. Immunoblot analysis was performed using TGF-β antibody (rabbit, 1 : 1000; Cell Signaling Technology (CST), USA), Smurf2 antibody (rabbit, 1 : 1000; Abcam, USA), actin antibody (rabbit, 1 : 1000; Abcam, USA), Smad7 antibody (rabbit, 1 : 500; Boster Biological Technology, China). Horseradish peroxidase-conjugated secondary antibodies (anti-rabbit) were obtained from the Beyotime Institute of Biotechnology, China. Proteins were detected using the enhanced chemiluminescence (ECL) system and ECL Hyperfilm (Millipore, USA).

The TGF-β antibody (#3709), TGF-β receptor II antibody (#41896), Phopho-SMAD2 (Ser465/Ser467) antibody (#18338), SMAD2 antibody (#5339), and cyclin D1 (#2978) antibody were obtained from Cell Signaling. The α-smooth muscle Actin (ab89989) antibody was purchased from Abcam.