All lanes : ab11173 diluted 1/5000Lane 1 : African Green Monkey COS-7 whole cell lysate with 5% MilkLane 2 : African Green Monkey COS-7 whole cell lysate with 5% MilkLysates/proteins at 20 µg per lane.SecondaryHRP conjugated Goat anti-rabbit polyclonal diluted 1/4000developed using the ECL techniquePerformed under reducing conditions.Observed band size : 200 kDa (why is the actual band size different from the predicted?)Exposure time : 20 secondsThis image is courtesy of an anonymous AbreviewThe gel running conditions were reducing.  The blocking time was 2 hours at 25°C.The primary antibody was incubated for 16 hours at 4°C.See Abreview

IHC image of ab11173 staining in human kidney 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 ab11173, 1µg/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.