C57BL/6 mice were sacrificed after 8 weeks of HFD feeding. Pancreata were removed, fixed in 10% formalin, and embedded in paraffin. The tissue sections were then incubated with primary antibody solution: guinea-pig anti-insulin (DAKO, 1:100) and rabbit anti-glucagon (DAKO, 1:100). Texas Red-conjugated goat anti-guinea-pig IgG (Santa Cruz Biotechnology, 1:200) and fluorescein isothiocyanate-conjugated goat anti-rabbit IgG (Santa Cruz Biotechnology,1:200) were used as secondary antibodies. Fluorescence was imaged using a laser scanning confocal fluorescent microscope (LSM 700, Carl Zeiss MicroImaging, Jena, Germany). The areas of alpha cells, beta cells, and islets were imaged and analyzed by CQ1spinning disk confocal systems (Yokogawa Electric Corporation) using CellVoyager software.
Fluorescein isothiocyanate conjugated goat anti rabbit igg
Manufactured by Santa Cruz Biotechnology
Fluorescein isothiocyanate-conjugated goat anti-rabbit IgG is a secondary antibody that binds to rabbit immunoglobulin G (IgG) and is conjugated to the fluorescent dye fluorescein isothiocyanate (FITC). This reagent is used in immunological detection and visualization techniques.
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2 protocols using fluorescein isothiocyanate conjugated goat anti rabbit igg
1
Quantitative Analysis of Pancreatic Islets
2
Claudin-4 Immunofluorescence in Lung Tissue
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Immunofluorescence staining for claudin-4 was performed as previously described (34 (link)). Lung tissue sections were deparaffinized, permeabilized using 0.1% Triton X-100, and blocked with 5% BSA before incubation with anti-claudin-4 (1:200, OriGene Technologies, Inc.) antibody overnight at 4°C. Fluorescein isothiocyanate-conjugated goat anti-rabbit IgG (1:200; Santa Cruz Biotechnology) was used as the secondary antibody for counterstaining. After washing in PBS and DAPI staining, slides were observed and images were captured using a fluorescence microscope (Leica DM 2500; Leica Microsystems GmbH, Wentzler, Germany).
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