ab17199 staining Thrombin in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 5% milk for 30 minutes at room temperature; antigen retrieval was by heat mediation in Tris pH 9.0. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. An undiluted Biotin-conjugated horse anti-mouse IgG polyclonal was used as the secondary antibody.

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IHC image of Thrombin staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab17199, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.