Anti mouse lc3b antibody

Clinical tissues and mouse tumors were fixed with 4% paraformaldehyde, dehydrated with different ethanol concentrations, embedded in paraffin, and then sliced (4 μm). Sections were dewaxed with xylene, rehydrated with ethanol of a gradient concentration, and rinsed with distilled water. Antigen repair was then performed with 0.01 M sodium citrate buffer (pH 6.0) for 30 min, treatment with 3% H₂O₂ for 5 min, and permeabilization with 0.1% TritionX-100 in PBS for 15 min. The primary antibodies, including anti-human MMP-12 antibody (1:300; MAB919; R&D Systems, USA) and anti-mouse LC3B antibody (1:200; 83506; Cell Signaling Technology, USA) were incubated overnight, and the tissue sections were incubated with secondary antibodies containing horseradish peroxidase for 45 min. The sections were stained with hematoxylin-eosin (HE) and made transparent with xylene after dehydration. According to staining intensity, sections were scored as 0=no staining, 1=mild staining, 2=moderate staining, and 3=intense staining. Immune response rate was recorded as 0 (0% immune response cells), 1 (<5%), 2 (5-50%), 3 (>50%). The final score was obtained by combining staining intensity with immune response rate. A score equal to or greater than 3 was considered positive staining.