The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Use a concentration of 1 - 5 µg/ml. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.
PSCA (Prostate Stem Cell antigen) is a cell surface antigen, which is overexpressed in ~40% of primary prostate cancers and in as many as 100% of metastatic ones. PSCA is also overexpressed in a majority of transitional cell and pancreatic carcinomas. Antibody directed against PSCA inhibits tumorigenesis, slows tumor growth, prolongs survival and prevents metastasis in a preclinical zenograft model indicating that PSCA may have utility as a prognostic marker and/or therapeutic target in prostate cancer.
IHC image of ab56338 staining in human normal kidney 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 EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab56338, 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.
Liu N et al. Simultaneous and combined detection of multiple tumor biomarkers for prostate cancer in human serum by suspension array technology. Biosens Bioelectron47:92-8 (2013).
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