Immunohistochemical stains were performed on 6 stage III and IV melanoma tissue microarrays (TMA) (Ekmekcioglu et al., 2016 (link); Iida et al., 2017 (link)). After heat mediated epitope retrieval at pH 8.0 for 20 min, the sections were incubated with mouse monoclonal SUZ12, Clone 3D10 (1:500, ThermoFisher) and mouse monoclonal EZH2, Clone AC22 (1:200, Cell signaling Technology). IHC staining was performed using a Leica Bond Max automated stainer (Leica Biosystems, Buffalo Grove, IL). The IHC reaction was performed using Leica Bond Polymer Refine detection kit (Leica Biosystems). Immunoreactive cells were visualized using diaminobenzidine (DAB) as chromogen followed by counterstaining with hematoxylin. All IHC slides were scanned using an Aperio AT Turbo (Leica Biosystems). The scoring was performed by a pathologist (RL) using direct microscope evaluation. EZH2 and SUZ12 expression was considered positive when nuclear staining was present on tumor cells and it was evaluated by H-score, which assesses the percentage of positive cells (0 to 100) multiplied by the intensity of staining (0 to 3+), with a total score ranging from 0 to 300. H-scores for EZH2 or SUZ12 in patients harboring an NRAS or BRAF mutation were compared using a t test.
Clone ac22
Manufactured by Cell Signaling Technology
Clone AC22 is a laboratory reagent for use in research applications. It is an antibody that recognizes a specific protein target. The core function of Clone AC22 is to bind and detect the target protein in various experimental techniques.
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Lab products found in correlation
2 protocols using clone ac22
1
Immunohistochemical Profiling of Melanoma
2
Immunohistochemical Profiling of Melanoma
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Immunohistochemical stains were performed on 6 stage III and IV melanoma tissue microarrays (TMA) (Ekmekcioglu et al., 2016 (link); Iida et al., 2017 (link)). After heat mediated epitope retrieval at pH 8.0 for 20 min, the sections were incubated with mouse monoclonal SUZ12, Clone 3D10 (1:500, ThermoFisher) and mouse monoclonal EZH2, Clone AC22 (1:200, Cell signaling Technology). IHC staining was performed using a Leica Bond Max automated stainer (Leica Biosystems, Buffalo Grove, IL). The IHC reaction was performed using Leica Bond Polymer Refine detection kit (Leica Biosystems). Immunoreactive cells were visualized using diaminobenzidine (DAB) as chromogen followed by counterstaining with hematoxylin. All IHC slides were scanned using an Aperio AT Turbo (Leica Biosystems). The scoring was performed by a pathologist (RL) using direct microscope evaluation. EZH2 and SUZ12 expression was considered positive when nuclear staining was present on tumor cells and it was evaluated by H-score, which assesses the percentage of positive cells (0 to 100) multiplied by the intensity of staining (0 to 3+), with a total score ranging from 0 to 300. H-scores for EZH2 or SUZ12 in patients harboring an NRAS or BRAF mutation were compared using a t test.
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