Validation of six commercial antibodies for detection of heterologous and endogenous TRPM8 ion channel expression

Abstract

TRPM8 is a non-selective cation channel expressed in primary sensory neurons and other tissues, including prostate and urothelium. Its participation in different physiological and pathological processes such as thermoregulation, pain, itch, inflammation and cancer has been widely de-scribed, making it a promising target for therapeutic approaches. The detection and quantification of TRPM8 seems crucial for advancing in the knowledge of the mechanisms underlying its role in these pathophysiological conditions. Antibody-based techniques are commonly used for protein detection and quantification, although their performance with many ion channels, including TRPM8, is suboptimal. Thus, the search for reliable antibodies is of utmost importance. In this study, we characterized the performance of six TRPM8 commercial antibodies in three immuno-detection techniques: western blot, immunocytochemistry and immunohistochemistry. Different outcomes were obtained for the tested antibodies; two of them proved to be successful detecting TRPM8 in the three approaches while, in the conditions tested, the other four were acceptable only for specific techniques. Considering our results, we offer some insight into the usefulness of these antibodies for detection of TRMP8 depending on the methodology of choice.