All lanes : Anti-LRPPRC antibody (ab21864) at 1/1000 dilutionLane 1 : Total protein extract from E. coli with ~50ng to 100ng of a negative control fusion protein with an irrelevant antigen at 20 ug Lane 2 : Total protein extract from E. coli with ~50ng to 500ng of the antigen fusion protein at 20 ugSecondaryRabbit anti-mouse IgG + IgM, (H+L) horseradish peroxidase conjugated at 1/5000 dilution
IHC image of ab21864 staining in Human liver 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 ab21864, 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.