IHC image of Histone H1 staining in a section of formalin-fixed paraffin-embedded [human normal colon]*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was then incubated with ab11080, 1/1000 dilution, for 15 mins at room temperature. A goat anti-mouse biotinylated secondary antibody (ab6788, 1/1000 dilution), was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, ab7403 at a 1/10000 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

The wells were coated with ab200699 at 1µg/ml at 50µl/well overnight at 4°C, followed by a 5% BSA blocking step for 2h RT. Human IgG3 (ab138703) was then added starting at 20 µg/ml and plasma/serum at 1:500 and gradually diluted 1:4, 50µl/well for 2h. Ab201248 was then added at 1:10,000 dilution, 50µl/well for 2h. A HRP-streptavidin (ab7403) was used at 1:10,000 dilution for 1h.