Notch1 primary antibody

Knee joints were harvested and fixed in 10% neutral buffered formalin for 3 days, decalcified in Formic Acid Bone Decalcifier (Immunocal, Decal Chemical Corp.) for 7–10 days, paraffin processed, and embedded for sectioning. Tissues were sectioned at 5 µm and stained with ABH/OG. IHC analyses were performed on sections using traditional antigen retrieval and colorimetric development methodologies with the following primary antibodies: SOX9 (Santa Cruz), COL2A1 (Thermo Scientific), COL10A1 (Quartett), COL1A1 (Abcam), COL3A1 (Abcam), MMP-13 (Thermo Scientific), IL6 (Abcam), and phosphorylated STAT3 (Cell Signaling). TUNEL cell death assay was performed on sections using the in situ Cell Death Detection Kit, Fluorescein (Roche) according to the manufacturer’s instructions. IF analysis and β-galactosidase staining was performed on frozen sections. Knee joints were harvested and fixed in 4% paraformaldehyde (PFA) for 2 hours at 4 °C and decalcified with 14% EDTA at 4 °C for 10 days. Tissues were washed in sucrose gradient, embedded with Tissue-TEK OCT medium, snap frozen in liquid nitrogen and sectioned at 10 µm using a Lecia CM 1850 cryotome. The NOTCH1 primary antibody (Santa Cruz) was used for IF analysis. Beta-galactosidase staining was performed as previously described (63 (link)).