Immunohistochemical studies were performed on breast cancer surgical specimens using the avidin-biotin-peroxidase method (DakoCytomation, Carpinteria, CA, USA) on formalin-fixed, paraffin-embedded tissues (FFPE). All of the sections were counterstained using hematoxylin. The product names and the dilutions of primary antibodies against the specific markers used in the study are available upon request.
Avidin biotin peroxidase method
Manufactured by Agilent Technologies
Sourced in United States, Spain
The Avidin-biotin-peroxidase method is a laboratory technique used to detect and localize specific proteins or antigens within a sample. It utilizes the high-affinity binding between avidin and biotin, coupled with a peroxidase enzyme, to amplify and visualize the target of interest. This method provides a sensitive and reliable approach for qualitative analysis.
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Lab products found in correlation
2 protocols using avidin biotin peroxidase method
1
Immunohistochemical Analysis of Breast Cancer
2
Quantifying Salmonella and Macrophages in Tissue
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Three samples of each collected tissue were sectioned at 5 mm and routinely processed with poly-l-lysine (Sigma-Aldrich, St. Louis, MO, USA), deparaffinized in xylene, and rehydrated through graded alcohols to distilled water. Hematoxylin and eosin staining was performed as previously described. 27 Immunohistochemistry (IHC) assays were performed with an anti-Salmonella rabbit polyclonal antibody with a specificity that was previously determined and a mouse monoclonal antibody specific for porcine macrophages (clone 4E9/11) (Biorad). The antibody reactions were visualized by a standard avidin-biotin peroxidase method (Dako, Barcelona, Spain) using diaminobenzidine (Sigma-Aldrich) as chromogen substrate. 42 To assess the specificity of each reaction, negative controls (reactions lacking the primary antibody) were performed in parallel for each section stained. After IHC labeling and image acquisition, the load of Salmonella was quantified in 10 images for each preparation using the IHC profiler plugin in ImageJ. 28 The IHC profiler algorithm, which estimates the number of positive pixels in each image, was used to enumerate Salmonella and macrophages. Neutrophils were enumerated by counting each cell type in 10 randomly selected fields (400Â field) from each cut processed.
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