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Validation of antibodies for the specific detection of human TRPA1.
Virk, H S; Rekas, M Z; Biddle, M S; Wright, A K A; Sousa, J; Weston, C A; Chachi, L; Roach, K M; Bradding, P.
Afiliación
  • Virk HS; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom. hsv4@leicester.ac.uk.
  • Rekas MZ; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Biddle MS; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Wright AKA; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Sousa J; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Weston CA; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Chachi L; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Roach KM; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
  • Bradding P; Department of Respiratory Sciences, University of Leicester, UK Institute of Lung Health and NIHR Leicester BRC-Respiratory, Leicester, United Kingdom.
Sci Rep ; 9(1): 18500, 2019 12 06.
Article en En | MEDLINE | ID: mdl-31811235
The transient receptor potential cation channel family member ankyrin 1 (TRPA1) is a potential target for several diseases, but detection of human TRPA1 (hTRPA1) protein in cells and tissues is problematic as rigorous antibody validation is lacking. We expressed hTRPA1 in a TRPA1-negative cell line to evaluate 5 commercially available antibodies by western blotting, immunofluorescence, immunocytochemistry and flow cytometry. The three most cited anti-TRPA1 antibodies lacked sensitivity and/or specificity, but two mouse monoclonal anti-TRPA1 antibodies detected hTRPA1 specifically in the above assays. This enabled the development of a flow cytometry assay, which demonstrated strong expression of TRPA1 in human lung myofibroblasts, human airway smooth muscle cells but not lung mast cells. The most cited anti-TRPA1 antibodies lack sensitivity and/or specificity for hTRPA1. We have identified two anti-TRPA1 antibodies which detect hTRPA1 specifically. Previously published data regarding human TRPA1 protein expression may need revisiting.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Miocitos del Músculo Liso / Miofibroblastos / Canal Catiónico TRPA1 / Anticuerpos Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Miocitos del Músculo Liso / Miofibroblastos / Canal Catiónico TRPA1 / Anticuerpos Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Reino Unido