This antibody labels melanocytes in the cytoplasm and plasma membrane and is useful tool for the identification of melanomas and clear cell sarcomas. It seems to be a melanocyte antibody of autoimmune origin as it does not react with the non-melanocytic peptide antigen used for immunization.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval with 1 mM EDTA buffer, pH 9.0 before commencing with IHC staining protocol. We suggest an incubation period of 30-60 minutes at room temperature. This antibody may be diluted to a titer of 1:10-1:25 in an ABC method.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre existing benign nevus, which occurs most often in the skin but also may involve other sites.
IHC image of ab12502 staining in human melanoma 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 ab12502, 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.