Validation of antibodies for the specific detection of human TRPA1
| Autor: | Adam K. A. Wright, Latifa Chachi, Katy M. Roach, Peter Bradding, J. Sousa, Michael Biddle, Cathryn Weston, Harvinder Virk, M. Z. Rekas |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
lcsh:Medicine Ion channels in the nervous system Epitopes 0302 clinical medicine Ankyrin lcsh:Science Myofibroblasts Lung TRPA1 Cation Channel chemistry.chemical_classification Multidisciplinary medicine.diagnostic_test biology Calcium signalling food and beverages Flow Cytometry Immunohistochemistry 3. Good health Blot Monoclonal Antibody psychological phenomena and processes Immunocytochemistry Green Fluorescent Proteins Myocytes Smooth Muscle Immunoblotting Immunofluorescence Antibodies Article Flow cytometry Target validation 03 medical and health sciences medicine Humans Calcium Signaling Gene Expression Profiling lcsh:R Molecular biology 030104 developmental biology HEK293 Cells chemistry Gene Expression Regulation Microscopy Fluorescence Cell culture biology.protein lcsh:Q Calcium Ion channel signalling 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-9 (2019) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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