1ml NCL-L-CK20-561

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Publication protocol

Formalin-fixed, paraffin-embedded tissues were deparaffinized in xylene, rehydrated through serial dilutions of alcohol and washed in phosphate–buffered saline (pH 7.2). After pre-treatment with the antigen retrieval solution at 95°C for 40 minutes (Table 1), endogenous peroxidase activity was blocked in 3% hydrogen peroxide. Slides were then incubated with primary antibody at room temperature (20-25°C) for 30 min (Table 1). Diaminobenzidine was used as the chromogen for the immunostaining. Finally, sections were counterstained with hematoxylin and mounted (Missaoui et al., 2018). Specific positive controls were used for each antibody. Negative controls were obtained by excluding the primary antibody. Images were captured by the microscopic digital camera Olympus system. Immunohistochemistry evaluation was independently performed by two pathologists. All immunostaining were scored as positive or negative as indicated in Table 1.

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