α sma antibody

Liver tissue sections (3 µm) were hydrated and inactivated with 3% H2O2. Incubate with goat serum at room temperature for 30 min. iNOS antibody (Abcam, Cambridge, UK, ab115819, dilution 1:200), IL-10 antibody (Bioss, Beijing, China, bs-0698R, 1:200), TGF-β antibody (Affinity, Cincinnati, US, AF5347, dilution 1:200), α-SMA antibody (Affinity, Cincinnati, US, AF1032, dilution 1:200), COL1α1 antibody (Affinity, Cincinnati, US, AF7001, dilution 1:200), MMP9 antibody (Affinity, Cincinnati, US, AF5228, dilution 1:200) or Caspase 3 antibody (Affinity, Cincinnati, US, AF6311, dilution 1:200) were incubated overnight at 4°C. The tablets were washed the next day, and the second antibody labeled with horseradish peroxidase was incubated at 37°C for 60 min. Sections were developed with DIAMinobenzidine (DAB) and stained with hematoxylin. The slides were rinsed with distilled water and air-dried, sealed and placed under a microscope for observation. Image J software was used to evaluate the mean optical density of immunohistochemical images.

The locations and expressions of target proteins α-SMA were investigated by immunohistochemical staining within 5-mm tissue slides. The slides were deparaffinized with xylene and rehydrated with ethanol. Antigen retrieval was performed by placing the slides in Tris/EDTA buffer (pH 9.0) before heating for 10 minutes. Then the slides were treated with endogenous peroxidase blocker for 10 minutes and normal goat serum (10%) was utilized for 30 minutes to block nonspecific binding sites. Subsequently, the slides were incubated with α-SMA antibody (Affinity, Cincinnati, OH, USA) at 4°C overnight. The primary antibody was visualized under the light microscope using the DAB method according to the kit instructions (Maixin Biotechnology, Fuzhou, China).