Anti mpo

The lung tissues of mice were incubated in 4% paraformaldehyde for fixation and paraffin embedding. After dewaxing in xylene, paraffin-embedded sections were dehydrated in graded ethanol. Incubation for 30 min with 0.3% H₂O₂ in PBS was followed by blocking with PBS containing 1% BSA for 1 h. Proteins of interest were recognized by incubation with anti-MPO (Cell Signaling, 1:1000) at 4°C overnight. After washing with TBST, sections were incubated at room temperature for 30 min with SignalStain Boost IHC Detection Reagent (CST, United States). SignalStain DAB (Cell Signaling Technology) was then applied to the sections to reveal the intensity of the staining. Sections were captured using an Olympus BX-51 microscope (Olympus, Japan).

Immunohistochemical staining to evaluate myeloperoxidase (MPO) was performed as described previously (33 (link)). Briefly, paraffin-embedded lung tissue sections were deparaffinized before antigen retrieval. The lung sections were incubated with a solution of 3% H₂O₂ in methanol for 15 min to block endogenous peroxidase. The slides were exposed to rabbit polyclonal antibody (anti-MPO, 1:100, Cell Signaling Technology, Danvers, MA, USA). The slides were then washed and incubated for 30 min with rat-specific horseradish peroxidase polymer anti-rabbit antibody (Nichirei Biosciences, Tokyo, Japan), and then horseradish peroxidase substrate was added for 3 min. The lung sections were then counterstained with hematoxylin.