Anti-Progesterone Receptor antibody (ab131486)

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Publication protocol

Expression of four markers was evaluated using standard methods for immunohistochemistry and commercial antibodies, including rabbit anti-ERα (Cat. #06-935; MilliporeSigma, St. Louis, MO [52]); rabbit anti-Ki67 (Cat. #RM-9106-S1; Thermo Fisher Scientific [53]), a marker of proliferation; rabbit anti-PR (Cat. #ab131486; Abcam, Cambridge, MA [54]); and rabbit anti-Wnt5a (Cat. #ab174963; Abcam [55]). See Table 1 for a summary of these antibodies. In brief, sections were deparaffinized, hydrated through a series of alcohols, microwaved in 10 mM citrate buffer (pH 6) for antigen retrieval, and treated with hydrogen peroxide to quench endogenous peroxidases. Nonspecific binding was blocked with 1% milk protein in 5% normal goal serum (Cell Signaling Technology, Danvers, MA). Sections were incubated with primary antibodies (see Table 1 for concentrations) at 4°C for 14 to 16 hours and washed and incubated with secondary antibody (goat anti-rabbit, Cat. #ab64256; Abcam [56]), followed by streptavidin peroxidase complex (Abcam, Cat# ab64269). Diaminobenzidene chromogen (Cat. #ab64238; Abcam) was used to visualize reactions. Sections were counterstained with Harris hematoxylin (Thermo Fisher Scientific). Each immunohistochemical run included a negative control in which the primary antibody was replaced with 5% normal goat serum.

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