AIMS/HYPOTHESIS: Adequate 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. METHODS: Commercially 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. RESULTS: All 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/ INTERPRETATION: Proper positive and negative controls are an important prerequisite for the evaluation of FFAR1 expression.
AIMS/HYPOTHESIS: Adequate 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. METHODS: Commercially 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. RESULTS: All 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 humanFFAR1. CONCLUSIONS/ INTERPRETATION: Proper positive and negative controls are an important prerequisite for the evaluation of FFAR1 expression.
Authors: Elisabeth Christiansen; Maria E Due-Hansen; Christian Urban; Manuel Grundmann; Johannes Schmidt; Steffen V F Hansen; Brian D Hudson; Mohamed Zaibi; Stine B Markussen; Ellen Hagesaether; Graeme Milligan; Michael A Cawthorne; Evi Kostenis; Matthias U Kassack; Trond Ulven Journal: J Med Chem Date: 2013-01-22 Impact factor: 7.446
Authors: Robert Wagner; Gabriele Kaiser; Felicia Gerst; Elisabeth Christiansen; Maria E Due-Hansen; Manuel Grundmann; Fausto Machicao; Andreas Peter; Evi Kostenis; Trond Ulven; Andreas Fritsche; Hans-Ulrich Häring; Susanne Ullrich Journal: Diabetes Date: 2013-02-01 Impact factor: 9.461
Authors: Sibylle Sabrautzki; Gabriele Kaiser; Gerhard K H Przemeck; Felicia Gerst; Estela Lorza-Gil; Madhura Panse; Tina Sartorius; Miriam Hoene; Susan Marschall; Hans-Ulrich Häring; Martin Hrabě de Angelis; Susanne Ullrich Journal: Mol Metab Date: 2017-07-18 Impact factor: 7.422