Abstract Reliable antibody performance is essential for cancer research, particularly when investigating disease-associated targets such as TROP2. Conventional antibody validation methods often fall short in addressing the complexity of human tissue expression and disease heterogeneity. To overcome these limitations, an enhanced validation framework was developed that integrates disease-relevant tissue profiling. This approach combines multiple validation steps, including knock out validation testing and generating detailed IHC expression profiles in selected formalin-fixed paraffin-embedded (FFPE) human tissue microarrays (TMA) using automated immunohistochemistry (IHC) platforms. As a case study, a recombinant rabbit monoclonal antibody (RabMab®) targeting TROP2, clone EPR20043 (ab214488) was validated across multiple IHC automated platforms. The antibody demonstrated high specificity, with no TROP2 detected by IHC in FFPE cell pellets prepared from TACSTD2 knockout MCF7 cells (ab286330). Furthermore, AI-driven expression profiling of multi-cancer TMAs (breast, lung, ovarian and endometrial cancer) revealed high sensitivity for detecting diverse protein expression levels, supporting its potential for biomarker assessment, patient stratification and diagnostic applications across varied disease settings. This study highlights the value of enhanced antibody validation in de-risking reagent selection and ensuring robust, reproducible results in cancer research. The TROP2 case exemplifies how rigorous Abcam’s antibody validation can accelerate discovery and translational applications in the oncology research. Citation Format: Caroline S. Hirst, JK Tanjore Ramanathan, Nadine Nelson, Antonella Galli, Will Howat, Silvia Sbacchi. Reliable antibodies for cancer research: TROP2 as a case study of Abcam's enhanced validation framework abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7733.
Hirst et al. (Fri,) studied this question.