Goat anti rabbit fluorescent secondary antibody

Heterotopic lipid formation was observed using haematoxylin eosin (HE) staining, and immunofluorescence for Perilipin 1 was used to observe the efficiency of hAMSC adipogenic differentiation in vivo. Briefly, paraffin sections with a thickness of 3 μm were prepared according to standard protocols, and then dewaxed in xylene, rehydrated through decreasing concentrations of ethanol, and washed in PBS. For HE staining, the sections were stained with haematoxylin and eosin. After staining, sections were dehydrated through increasing concentrations of ethanol and xylene and mounted. For immunofluorescence, tissue antigens were retrieved by citric acid buffer (PH6.0) microwave antigen retrieval (Servicebio, Wuhan, China) at first. Then, the sections were blocked for 30 min at room temperature in 10% BSA (Servicebio), perilipin 1 was detected by incubation with anti- perilipin 1 antibody (Cell Signalling Technology) at 4°C overnight. Sections were rinsed three times in PBS and incubated with Goat anti-Rabbit fluorescent secondary antibody (Servicebio) for 1 h at room temperature the next day, and then cell nucleus were stained with DAPI (Servicebio) for 5 mins at room temperature. After rinsing three times in PBS, expression of perilipin 1 protein was observed under fluorescence microscope.