IHC image of Smad4 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab195554 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. 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
![Anti-Smad4 antibody [EP618Y] (HRP) (ab195554) at 1/5000 dilution + SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µgdeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/1947_ab195554-238556-WBab1955541.jpg)
Anti-Smad4 antibody [EP618Y] (HRP) (ab195554) at 1/5000 dilution + SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µgdeveloped using the ECL techniquePerformed under reducing conditions.