IHC image of NIPA staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab63557, 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.
All lanes : Anti-NIPA (phospho S354) antibody (ab63557) at 1/500 dilutionLane 1 : extracts from COS7 cells, treated with HU (2nM, 24hours)Lane 2 : extracts from COS7 cells, treated with HU (2nM, 24hours) with immunizing phosphopeptide at 10 µgLysates/proteins at 30 µg per lane.