Rabbit anti aldh1a1 polyclonal antibody

The paraffin-embedded sections were cut at a thickness of 2 μm. The immunohistochemical analysis was performed as previously described 20 . After being baked at 60 °C for 60 minutes, the sections were dewaxed and rehydrated with xylene and graded alcohol. After antigen retrieval, the sections were incubated overnight at 4°C with primary antibodies as follows: rabbit anti-CCR9 polyclonal antibody (1:200, Abcam), rabbit anti-CCL25 polyclonal antibody (1:200, ABclonal and 1:200, Signalway Antibody), rabbit anti-ALDH1A1 polyclonal antibody (1:400,Abcam). The sections were washed with PBS and developed with diaminobenzidine (DAB). Finally, the sections were counterstained with hematoxylin, dehydrated and then sealed.

Immunohistochemistry was routinely conducted as previously described [27 (link), 28 (link)]. After antigen retrieval, the slides were incubated with the following primary antibodies: NSE (1:100; Abcam), rabbit anti-ALDH1A1 polyclonal antibody (1:200; Abcam), and NBL1 (1:200; Affinity) overnight at 4 °C. Immunohistochemical staining was performed using diaminobenzidine (DAB). After counterstaining with hematoxylin, the sections were dehydrated and sealed. Staining scores were assessed by two different individuals (ZQZ and PLZ). The percentage and intensity of staining were evaluated and multiplied to obtain the final scores. The principle of immunohistochemical scoring is mainly based on staining intensity (0–3 points) and staining area (0–3 points). Finally, the product is the total score. Dyeing intensity was as follows: 0 points for no dyeing, 1 point for light yellow, 2 points for yellow or brown, 3 points for brown or tan; the stained area was as follows: 0 points for ≤5%, 1 point for 5–25%, 2 points for 25–50%, and 3 points for ≥50%. An immunohistochemical score of ≤3 indicated a low expression group, and a score >3 indicated a high expression group [29 (link)].