ab47153 staining HeLa cells.ICC/IF was performed with a standard paraformaldehyde technique (non permeabilized, blocked with 0.2% fish gelatin for 30 min at 25°C). Primary antibody used at 1/200 in 0.12% PBS / fish gelatin for one hour. The secondary antibody was a Cy3-conjugated Goat anti-Mouse. See Abreview
IHC image of ab47153 staining in human tonsil 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 ab47153, 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.
A typical staining pattern with the B-L2 monoclonal antibody of lymphocytes