Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues.

Antibodies are key reagents in cell biology and biochemistry research. The validation of their performance, in terms of sensitivity and specificity, is essential for their correct application. TRPV1 and TRPA1 are non-selective cation channels expressed in primary sensory neurons, where they mediate the detection of diverse physical and chemical stimuli. They play key roles in nociception and inflammatory processes, making them important targets for mechanistic and therapeutic pain studies, highlighting the need for reliable evaluation of their expression. The detection and quantification of TRPV1 and TRPA1 protein expression is commonly carried out using antibody-based techniques, such as immunohistochemistry and western blotting. However, as with other TRP channels and membrane proteins, antibody performance is frequently suboptimal, leading to potential misinterpretation of results and erroneous conclusions. In this study, we systematically evaluated the performance of five TRPV1 and seven TRPA1 commercial antibodies in immunocytochemistry, immunohistochemistry, and western blotting, using both heterologous and native expression systems. We identified two TRPV1 antibodies that consistently yielded robust and specific signals across all techniques and expression models tested; their specificity was validated using a TRPV1 KO mouse. For the remaining antibodies, we provide guidance to facilitate the selection of the most appropriate reagent according to the experimental approach.