Scientist Abcam CAMBRIDGE, England, United Kingdom
Disclosure(s):
JK Ramanathan, PhD: No financial relationships to disclose
Introduction/Rationale: Reliable antibody performance is essential for translational research, particularly when investigating disease-associated targets such as, Triggering Receptor Expressed on Myeloid Cells 1 (TREM1), a key modulator of innate immune responses implicated in inflammatory conditions. Conventional antibody validation methods often fall short in addressing the complexity of human tissue expression and disease heterogeneity.
Methods: To overcome these limitations, an enhanced validation framework was developed that integrates disease-relevant tissue profiling. This approach combines multiple validation steps, including specificity 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 TREM1, clone EPR22060-229 (ab225861), was validated across multiple IHC automated platforms.
Results: In both normal and cancer tissues, the antibody showed positive staining in interstitial regions and infiltrating lymphocytes. Its high specificity was further validated through co-localization assays with CD14, a well-established myeloid marker, across COVID-19, glioblastoma, and inflammatory bowel disease (IBD) samples, confirming accurate detection of TREM1-expressing cells.
Conclusion: This study highlights the value of enhanced antibody validation in de-risking reagent selection and ensuring robust, reproducible results in translational research. The TREM1 case exemplifies how rigorous Abcam’s antibody validation can accelerate discovery and translational applications.
