OSTEOPONTIN (O-17) ANTI-HUMAN RABBIT IGG AFFINITY PURIFY

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Fixed hearts were embedded in paraffin and sectioned at 5 μm. Antigen retrieval was performed in 10 mmol/L citrate buffer antigen, endogenous peroxidase activity was blocked by immersing the sections in 3% H2O2, and nonspecific epitope binding was blocked by incubation with 4% bovine serum albumin. The sections were then incubated with NOX4 polyclonal antibody (Santa Cruz Biotechnology, USA) and OPN polyclonal antibody (IBL, Japan) overnight at 4 °C, followed by an incubation for 30 min at room temperature with horse-radish peroxidase (HRP)-conjugated secondary antibody (1:200), and visualized with diaminobenzidine.

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