The tissues of 24-week-old mice were fixed with 4% paraformaldehyde. Frozen sections of the liver were stained with Oil red O. Paraffin-embedded sections of the adipose tissue and liver were stained with hematoxylin and eosin, Masson’s trichrome, or Sirius red. More than 200 cells were analyzed for determination of the average adipocyte size using ImageJ software (National Institutes of Health). Masson’s trichrome– or Sirius red–positive areas were used to estimate the extent of fibrosis. For immunofluorescence, sections were incubated with Mac-2 antibody (Cedarlane) and perilipin antibody (Sigma-Aldrich) overnight. The sections were incubated with Alexa Fluor 488 (Thermo Fisher Scientific) for Mac-2 and/or Alexa Fluor 568 (Thermo Fisher Scientific) for perilipin. Nucleus was stained with 4′,6-diamidino-2-phenylindole (DAPI). Images of sections were captured using a confocal laser microscope (FV1000; Olympus). Perilipin staining was used to identify necrotic adipocytes. More than 400 cells were analyzed for determination of the necrotic adipocytes.
Perilipin antibody
Manufactured by Merck Group
Perilipin antibody is a laboratory reagent used to detect and quantify the expression of the perilipin protein. Perilipin is an intracellular lipid droplet-associated protein that plays a role in the regulation of lipolysis. The antibody can be used in various techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of perilipin in biological samples.
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2 protocols using perilipin antibody
1
Histological Analyses of Adipose and Liver
2
Quantifying Bone-Adipose Interactions
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The 4% PFA‐fixed, 18% EDTA‐decalcified, and paraffin‐embedded left femur samples were prepared and sectioned into 5 µm‐thick slices. Immunofluorescence staining for perilipin was conducted to test the changes of bone marrow adipocytes. Perilipin antibody was purchased from Sigma‐Aldrich. Osteogenic and osteoclastic activities, respectively, were evaluated by immunohistochemical staining for OCN using antibodies from Servicebio (Wuhan, China) and TRAP staining using a kit from Sigma‐Aldrich. Adipocytes, OCN+ osteoblasts, and TRAP+ osteoclasts were counted from four random visual fields of distal metaphysis for each femur section and five mouse femur samples for each group. The numbers of adipocytes per area (N. AdCs/Ar/mm2) and the numbers of osteoblasts (N. OBs/BS/mm) and osteoclasts (N. OCs/BS/mm) per BS were calculated. Immunofluorescence double staining for DMP1/NPY and immunohistochemical staining for NPY were conducted to assess DMP1 and NPY expression in the bone tissues. Immunofluorescence double staining for DMP1/TH and DMP1/VaChT was performed to evaluate the ANS innervation of osteocytes. Anti‐NPY and anti‐TH were obtained from Cell Signaling Technology (Danvers, USA). Anti‐DMP1 and anti‐VaChT were purchased from Novus Biologicals (Littleton, USA) and Sigma‐Aldrich, respectively.
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