Anti ly6g antibody

Paraffin-embedded kidney slides were processed sequentially through deparaffinization, rehydration, and antigen retrieval. Subsequently, the slides were subjected to a 15-minute incubation with a hydrogen peroxide solution at room temperature, followed by a 1-hour blocking step with 10% goat serum. Subsequently, the slices were incubated overnight at 4 °C with the primary antibody. The following day, the sections were allowed to equilibrate at room temperature for at least 30 min. Subsequently, the sections were treated with diaminobenzidine substrate and counterstained with hematoxylin before being mounted. Finally, the slices were examined under a Nikon microscope (Japan). Three kidney specimens from all groups were blindly caught across three equal microscopic fields (400x) for analysis. With the purpose of reckoning average optical density (AOD), we operated integrated optical density (IOD) measurements on IHC pictures. AOD = IOD / area. After delimiting the zone of interest, the AOD of the chosen precinct (IOD per unit area) was determined utilizing Image-J software. AOD depicts the protein immunoreactivity in the kidneys. The primary antibody used in this experiment was Anti-LY6G antibody (1:100, Lot No: ab25377) (Abcam, USA).

Histological analyses were performed essentially as described before. Briefly, the paraffin embedded sections were blocked with 10% normal goat serum for 1 h at room temperature and then incubated with an anti‐Ly6G antibody (Abcam, cat# ab238132, 1:100) or anti‐BRG1 antibody (Abcam, cat# ab110641, 1:100). Staining was visualized by incubation with anti‐rabbit secondary antibody and developed with a streptavidin‐horseradish peroxidase kit (Pierce, cat# 21140) for 20 min. Pictures were taken using an Olympus IX‐70 microscope. Slides were observed under a light microscope at high power (X40) by two pathologists independently in a double‐blind fashion. The scoring system was based on the following criterion: the staining intensity was divided into quintiles; the slides with the strongest staining were given a score of 5 (top quintile) whereas the slides with the dimmest staining were given a score of 1 (bottom quintile).