IF was performed as described previously 82 (link),83 (link). Briefly, after antigen retrieval using citrate buffer (Sigma), 5μm-thick formalin-fixed pancreatic sections were incubated with anti-insulin (1:1000, Agilent Cat#A0564, RRID:AB_10013624) or anti-glucagon primary antibodies (1:1000, Sigma-Aldrich Cat#G2654, RRID:AB_259852) and AlexaFluor-594-conjugated goat anti guinea-pig (1:500, Molecular Probes Cat#A-11076, RRID:AB_141930) and AlexaFluor-488-conjugated goat anti-mouse antibodies (1:500, Thermo Fisher Scientific Cat#A-11001, RRID:AB_2534069). Images were processed for morphometry using ImageJ software by an observer blind to experimental groups.
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Immunofluorescence Imaging of Pancreatic Cells
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Corresponding Organization :
Other organizations : Université de Lille, Centre de Recherche Jean Pierre Aubert, Instituto de Investigación Sanitaria de Santiago, Universidade de Santiago de Compostela, Institut des Neurosciences Cellulaires et Intégratives, Institut Cochin, Saarland University, KU Leuven, VIB-KU Leuven Center for Cancer Biology, Cisbio (France), VIB-UGent Center for Medical Biotechnology, Beth Israel Deaconess Medical Center, Institut Pasteur de Lille
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Variable analysis
independent variables
- Antigen retrieval using citrate buffer (Sigma)
- Insulin expression
- Glucagon expression
- Formalin-fixed pancreatic sections
- Thickness of sections (5 μm)
- Positive control: Anti-insulin primary antibody (1:1000, Agilent Cat#A0564, RRID:AB_10013624)
- Positive control: Anti-glucagon primary antibody (1:1000, Sigma-Aldrich Cat#G2654, RRID:AB_259852)
- Negative control: Not explicitly mentioned
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