Antibodies for HSP90B1 (ab238126) were purchased from Abcam company. Samples were embedded in paraffin at a thickness of 4 μm. Deparaffinization and rehydration were performed on each slide. To eliminate aldehyde linkages from antigens, they were reextracted using a pressure cooker and 0.01 M citrate buffer (pH 6). The slides were incubated with HSP90B1 antibody (1: 2000) overnight. After incubating the HRP-labelled secondary antibody for 1 h, immunodetection was performed the following day using diaminobenzidine, according to the manufacturer’s instructions [61 (link)].
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