Our Abpromise guarantee covers the use of ab71495 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.5 - 4 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).|
|IP||Use a concentration of 2 - 5 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
Immunohistochemical analysis of formaldehyde-fixed paraffin-embedded murine obsese nephric tissue sections, labelling IL18 with ab71495 at a dilution of 1/2500 incubated for 12 hours at 4°C in PBS and 2.5% horse serum diluent. Heat mediated antigen retrival was with citrate buffer and blocking was with 2.5% horse serum in PBS incubated for 1 hour at 25°C. Secondary was a horse anti-rabbit polyclonal HRP conjugate.
IHC image of ab71495 staining in mouse normal spleen 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 ab71495, 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.