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Detection of free fatty acid receptor 1 expression: the critical role of negative and positive controls |
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| Authors: | Charlott-Amelie Teutsch Madhura Panse Manuel Grundmann Gabriele Kaiser Evi Kostenis Hans-Ulrich Häring Susanne Ullrich |
| Affiliation: | 1. Division of Endocrinology, Diabetology, Vascular Medicine, Nephrology and Clinical Chemistry, Department of Internal Medicine IV, University of Tübingen, Otfried-Müller-Str. 10, 72076, Tübingen, Germany 2. Institute of Pharmaceutical Biology, University of Bonn, Bonn, Germany 3. Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Center Munich at the University of Tübingen (IDM), Partner in the German Center for Diabetes Research (DZD), Tübingen, Germany |
| Abstract: | Aims/hypothesisAdequate evaluation of protein expression is a crucial prerequisite for functional studies. Commonly used strategies comprise detection of proteins by specific antibodies using western blotting and immunohistochemical staining, or detection of mRNA by in situ hybridisation and RT-PCR. We evaluated the tools for the detection of free fatty acid receptor 1 (FFAR1) expression.MethodsCommercially available antibody preparations were used to detect endogenous expression of the FFAR1 receptor and this was compared with cell preparations deficient or overexpressing the mouse or human receptor. Concentrations of mRNA were evaluated by RT-PCR.ResultsAll insulin-secreting cells, INS-1E, Min6 and mouse islets showed specific expression of Ffar1 at the mRNA level. However, none of the commercially available antibodies specifically detected rat, mouse or human FFAR1.Conclusions/interpretationProper positive and negative controls are an important prerequisite for the evaluation of FFAR1 expression. |
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