VENTANA anti-MSH2 (G219-1129) Mouse Monoclonal Primary Antibody

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Publication protocol

MMR protein IHC was performed on the Ventana Benchmark autostaining system utilizing primary antibodies from Ventana (Roche/Ventana Medical Systems, Tucson, AZ). MSH2 IHC used a mouse monoclonal antibody (clone G219-1129), MSH6 IHC used a mouse monoclonal antibody (clone 44), MLH1 IHC used a mouse monoclonal antibody (clone M1) and PMS2 IHC used a rabbit monoclonal antibody (clone EPR3947). All samples were incubated with primary antibody after antigen retrieval in CC1 buffer, and primary antibody incubation was followed by detection with the UltraView HRP system (Roche/Ventana Medical Systems, Tucson, AZ). Each tissue microarray spot or standard histologic section containing tumor cells was visually dichotomously scored for presence or absence of cytoplasmic MMR protein signal by a urologic pathologist blinded to the sequencing/MSI testing data (TLL). A spot was considered to show MMR protein loss if any tumor cells in any tumor spot showed MMR protein loss, with intact staining in admixed benign prostate glands and/or surrounding stromal cells, endothelial cells or lymphocytes. Spots without internal control staining were considered ambiguous and not scored. All samples were initially screened for MSH2 loss by scoring TMA spots; however for all cases with MSH2 loss on TMA, confirmatory immunostaining for MSH2, MSH6, MLH1 and PMS2 was also performed on standard histologic tissue sections.

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