Dia clone h09

The presence and extent of tumor was assessed on H&E slides. Immunohistochemical staining for mutant IDH1 was performed using the anti-human IDH1 R132H mouse monoclonal antibody DIA clone H09 (Dianova, Germany) at a dilution of 1:100. Pretreatment of slides with Cell Conditioner 1 (EDTA) MILD using the Ventana BenchMark XT stainer system was followed by incubation with the primary antibody for 32 minutes at 37°C, which was then detected using Ventana UltraView detection with Ventana DAB. Hematoxylin counterstain was performed with the Leica ST 5020 apparatus. Positive and negative controls were performed using specimens previously validated.

Tumor specimens were fixed in formalin and processed for paraffin embedding. Hematoxylin and eosin staining and immunohistochemistry were conducted on 5-micron-thick formalin-fixed, paraffin-embedded tissue sections mounted on Leica Surgipath slides followed by drying for 60 min at 70 °C. Immunohistochemistry with the anti-IDH1-R132H antibody (Monoclonal Mouse Anti-human IDH1 (R132H), Dianova, DIA Clone H09) and DAB chromogen was performed on a Leica Bond III instrument using Bond Polymer Refine Detection System (Leica Microsystems AR9800) following a 20 min heat-induced epitope retrieval with Epitope Retrieval 2, EDTA, pH 9.0 [23 (link)].