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Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

TRPM8 is a non-selective cation channel expressed in primary sensory neurons and other tissues, including the prostate and urothelium. Its participation in different physiological and pathological processes such as thermoregulation, pain, itch, inflammation and cancer has been widely described, maki...

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Autores principales: Hernández-Ortego, Pablo, Torres-Montero, Remedios, de la Peña, Elvira, Viana, Félix, Fernández-Trillo, Jorge
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784522/
https://www.ncbi.nlm.nih.gov/pubmed/36555804
http://dx.doi.org/10.3390/ijms232416164
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author Hernández-Ortego, Pablo
Torres-Montero, Remedios
de la Peña, Elvira
Viana, Félix
Fernández-Trillo, Jorge
author_facet Hernández-Ortego, Pablo
Torres-Montero, Remedios
de la Peña, Elvira
Viana, Félix
Fernández-Trillo, Jorge
author_sort Hernández-Ortego, Pablo
collection PubMed
description TRPM8 is a non-selective cation channel expressed in primary sensory neurons and other tissues, including the prostate and urothelium. Its participation in different physiological and pathological processes such as thermoregulation, pain, itch, inflammation and cancer has been widely described, making it a promising target for therapeutic approaches. The detection and quantification of TRPM8 seems crucial for advancing 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 immunodetection techniques: Western blot, immunocytochemistry and immunohistochemistry. Different outcomes were obtained for the tested antibodies; two of them proved to be successful in 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 the detection of TRPM8 depending on the methodology of choice.
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spelling pubmed-97845222022-12-24 Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression Hernández-Ortego, Pablo Torres-Montero, Remedios de la Peña, Elvira Viana, Félix Fernández-Trillo, Jorge Int J Mol Sci Article TRPM8 is a non-selective cation channel expressed in primary sensory neurons and other tissues, including the prostate and urothelium. Its participation in different physiological and pathological processes such as thermoregulation, pain, itch, inflammation and cancer has been widely described, making it a promising target for therapeutic approaches. The detection and quantification of TRPM8 seems crucial for advancing 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 immunodetection techniques: Western blot, immunocytochemistry and immunohistochemistry. Different outcomes were obtained for the tested antibodies; two of them proved to be successful in 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 the detection of TRPM8 depending on the methodology of choice. MDPI 2022-12-18 /pmc/articles/PMC9784522/ /pubmed/36555804 http://dx.doi.org/10.3390/ijms232416164 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hernández-Ortego, Pablo
Torres-Montero, Remedios
de la Peña, Elvira
Viana, Félix
Fernández-Trillo, Jorge
Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title_full Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title_fullStr Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title_full_unstemmed Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title_short Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression
title_sort validation of six commercial antibodies for the detection of heterologous and endogenous trpm8 ion channel expression
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784522/
https://www.ncbi.nlm.nih.gov/pubmed/36555804
http://dx.doi.org/10.3390/ijms232416164
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