ab50591 staining Actin in mouse proximal colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% NDS for 24 hours at 4°C; antigen retrieval was by heat mediation in buffer, pH9. Samples were incubated with primary antibody (1/100 in 5% BSA/NDS) for 24 hours at 4°C. An undiluted Alexa Fluor^® 647-conjugated donkey anti-rat IgG polyclonal was used as the secondary antibody.

Also stained with ab76055 (green) at 1/100.

See Abreview

IHC image of Actin staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab50591, 5µg/ml, for 15 mins at room temperature. A Goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.