IHC image of ab68735 staining in human hippocampus 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 ab68735, 5µg/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-NSMase2 antibody (ab68735) at 1/500 dilutionLane 1 : nSMase2 transfected 293T cell lysateLane 2 : Non transfected 293T cell lysateLysates/proteins at 25 µg per lane.SecondaryGoat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution