Immunohistochemical staining kit

Rats without BALF collection were selected for lung histopathological evaluation. Rats were handled 24 h after LPS inhalation, taken out the lung tissues and washed with PBS to prepare sections. Hematoxylin Eosin (HE) staining: dewaxed and stained with hematoxylin for 5 min, returned to blue in 0.6% ammonia, rinsed and stained 3 min, dehydrated with ethanol, and sealed with neutral gum for observation and analysis. Isotype controls were processed without primary antibody incubation. Lung pathological changes was examined under a microscope (Eclipse 80i, Nikon, Japan) at 20x magnification. Immunohistochemical staining: Anti-Socs3 primary antibody (1:200, Abcam, UK) was added after the sections were deparaffinized and incubated at 37 °C for 1 h. Secondary antibodies and DAB staining were incubated according to the immunohistochemical staining kit (Maixin Biotechnology Co., Ltd., China) instructions. Then, lung sections were counterstained with hematoxylin. Staining was visualized under a microscope at 20x magnification. And the positive staining area was analyzed with Image J software.

Paraffin-embedded tissue samples were serially sectioned and immunohistochemically examined using an immunohistochemical staining kit (Maixin Bio, China), according to the manufacturer’s instructions, with antibodies against Ki-67 (Maixin Bio, China).