Sox-9 Antibody (E-9): sc-166505

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Pathologically confirmed chordoma tissues were obtained from patients who had undergone surgical resection in Massachusetts General Hospital, and were conducted according to the policies of the institutional review board of the hospital (IRB protocol # 2007P-002464). A retrospective study that included 50 samples from 50 chordoma patients was identified for the tissue TMA immunohistochemical staining. The expression level of SOX9 was determined based on the Immunohistochemistry Protocol (Paraffin) from Cell Signaling Technology (Danvers, MA). Briefly, each paraffin embedded slide was baked at 60°C for 1 hour. Each section was washed three times in xylene for 5 minutes, and then transferred through graded ethanol (100% and 95%) twice for 10 minutes. Following the process of antigen retrieval, endogenous peroxidase activity was quenched by incubation in 3% hydrogen peroxide. After protein blocking with normal goat serum for 1 hour at room temperature, the SOX9 primary antibody (Sox-9 (H-90): sc-20095, Santa Cruz Biotechnology, 1:50 dilution) was applied at 4°C overnight in a humidified chamber. The bound antibody on the array was detected by using SignalStain®Boost Detection Reagent and SignalStain® DAB (Cell Signaling Technology). Prior to imaging, the section was counterstained with hematoxylin QS (Vector Laboratories) and mounted with VectaMount AQ (Vector Laboratories) coverslip for long-term preservation.

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