Anti-MMP3 antibody (ab53015)

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DA rats cartilage tissues were fixed in 4% buffered paraformaldehyde for more than 2 days and subsequently decalcified with EDTA (12.5% EDTA, pH 7.4). Cartilage tissue sections were embedded in paraffin and 4 μm-thick sections were cut. Immunohistochemistry staining was performed using the standard streptavidin peroxidase complex method. Firstly, tissue sections were deparaffinized in xylene and rehydrated through a graded series of alcohols. Next, tissue sections were subjected to H2O2 treatment and antigen retrieval, and subsequently blocked with 5% bovine serum albumin (BSA) for 30 min at room temperature. Tissue sections were incubated with primary antibodies at 4°C overnight. After sequential incubations with biotinylated secondary antibody (BOSTER, catalog SA1022, China) and horseradish peroxidase-conjugated avidin (BOSTER, catalog SA1022, China), protein staining was performed using the Diaminobenzidine Substrate Kit (BOSTER, catalog SA2022, China). Samples were counterstained with hematoxylin, and the resulting brown staining indicated the proteins immunoreactivity. The results of IHC were observed under microscope with 400× magnification and quantification was performed with the Image-Pro® Plus software. The antibody information is depicted in Table S3.

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