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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
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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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
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
Zeitschrift
Diabetologia
Quellenangaben
Band: 57,
Heft: 4,
Seiten: 776-780
Verlag
Springer
Verlagsort
Berlin ; Heidelberg [u.a.]
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed