Anti sox11

After sectioning, the tissues were incubated with 3% H₂O₂ solution for 10 min, washed, and trypsinized for 5 min to unmask target antigens. Slides were then incubated with blocking solution [4% bovine serum albumin (BSA)] for 2 h at room temperature and incubated with primary antibodies (anti-SOX11, anti-HER2, or anti-ALDH1 antibodies; Abcam) at 4 °C for 8 h. Slides were then washed three times in 1× phosphate buffered saline (PBS) (2 min each), followed by treatment with secondary antibody at room temperature for 45 min. After washing, the slides were overlaid with a freshly prepared DAB solution, incubated at room temperature for 5 min, washed once in 1× PBS, and counterstained with hematoxylin.

Dissected tissue samples were mechanically lysed in ice-cold sample buffer. Cell lysates were then boiled for 10 min and stored at −80 °C until needed. Samples were loaded in equal concentrations (60 µg), resolved on 12% SDS-PAGE gel run at 50 V for 4 h, transferred to a PVDF membrane, and blocked in 5% non-fat milk for 1 h. The membrane was then incubated in primary antibody solution (anti-SOX11, anti-HER2, or anti-ALDH1 antibody; Abcam) at 4 °C overnight with gentle agitation. Blots were then washed, incubated with secondary antibody, and visualized with DAB solution.