IHC image of ab59497 staining in human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab59497, 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.
All lanes : Anti-P4HA1 antibody (ab59497) at 1 µg/mlLane 1 : Whole cell lysate prepared from 9L cellsLane 2 : Whole cell lysate prepared from PC12 cellsLane 3 : Whole cell lysate prepared from RIE-1 cellsLysates/proteins at 20 µg per lane.SecondaryHRP conjugated rabbit anti-goat polyclonal at 1/5000 dilutiondeveloped using the ECL technique
Anti-P4HA1 antibody (ab59497) at 0.2 µg/ml + HEK293 cell lysate (35µg protein in RIPA buffer)