Detection of free fatty acid receptor 1 expression: the critical role of negative and positive controls

作者：Teutsch Charlott Amelie; Panse Madhura; Grundmann Manuel; Kaiser Gabriele; Kostenis Evi; Haering Hans Ulrich; Ullrich Susanne^*

来源：Diabetologia, 2014, 57(4): 776-780.

DOI：10.1007/s00125-014-3161-8

• Antibodies
• FFAR1/GPR40
• Ffar1expression
• RT-PCR
• Western blotting

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 mRNAwere 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.

• 出版日期2014-4