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Open Access Green as soon as Postprint is submitted to ZB.
Detection of free fatty acid receptor 1 expression: The critical role of negative and positive controls.
Diabetologia 57, 776-780 (2014)
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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Antibodies ; Ffar1/gpr40 ; Ffar1expression ; Rt-pcr ; Western Blotting; Pancreatic Beta-cells; Insulin-secretion; Redistribution; Gpr40; Stimulation
ISSN (print) / ISBN
0012-186X
e-ISSN
1432-0428
Journal
Diabetologia
Quellenangaben
Volume: 57,
Issue: 4,
Pages: 776-780
Publisher
Springer
Publishing Place
Berlin ; Heidelberg [u.a.]
Reviewing status
Peer reviewed