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.
Anti perilipin 1 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-Perilipin-1 antibody is a laboratory reagent used to detect and quantify the Perilipin-1 protein in samples. Perilipin-1 is a lipid droplet-associated protein that plays a role in the regulation of lipolysis. This antibody can be used in various applications, such as Western blotting, immunohistochemistry, and immunofluorescence, to study the expression and localization of Perilipin-1 in cells and tissues.
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3 protocols using anti perilipin 1 antibody
1
Analyzing Adipogenic Differentiation of hAMSCs
2
Immunohistochemical Analysis of Adipocyte Proliferation
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Immunohistochemical analysis was performed with a rabbit polyclonal anti-Perilipin-1 antibody (Cell Signaling Technology, MA, USA) for proliferation of adipocytes. On the frozen specimens of the gastrocnemius muscle, endogenous peroxidase was inactivated by 0.3% hydrogen peroxide in methanol for 30 minutes. The sections were incubated with the primary antibody overnight at 4 °C. Between the incubation steps, sections were dip-immersion washed (3 ⋅ 5-minute wash) in tris-buffered saline (TBS) to eliminate excesses of non-bound antibodies or reagent. The antibody was diluted in 1% BSA/TBS to suppress nonspeci c reactions. Then, the sections were incubated with the anti-rabbit immunoglobulin conjugate horseradish peroxidase (HRP) and anti-mouse immunoglobulin conjugate HRP mixed in by employing the universal immuno-enzyme polymer method (Histo ne® Simple Stain MAX-PO; Nichirei, Tokyo, Japan). The reaction products were visualized in 0.15 mg/ml 3,3'-diaminobenzidine tetrahydrochloride (DAB) solution containing 0.003% hydrogen peroxide. After washing in water, the counter was stained by hematoxylin.
3
Immunohistochemical Analysis of Adipocyte Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical analysis was performed with a rabbit polyclonal anti-Perilipin-1 antibody (Cell Signaling Technology, MA, USA) for proliferation of adipocytes. On the frozen specimens of the gastrocnemius muscle, endogenous peroxidase was inactivated by 0.3% hydrogen peroxide in methanol for 30 minutes. The sections were incubated with the primary antibody overnight at 4 °C. Between the incubation steps, sections were dip-immersion washed (3 ⋅ 5-minute wash) in tris-buffered saline (TBS) to eliminate excesses of non-bound antibodies or reagent. The antibody was diluted in 1% BSA/TBS to suppress nonspeci c reactions. Then, the sections were incubated with the anti-rabbit immunoglobulin conjugate horseradish peroxidase (HRP) and anti-mouse immunoglobulin conjugate HRP mixed in by employing the universal immuno-enzyme polymer method (Histo ne® Simple Stain MAX-PO; Nichirei, Tokyo, Japan). The reaction products were visualized in 0.15 mg/ml 3,3'-diaminobenzidine tetrahydrochloride (DAB) solution containing 0.003% hydrogen peroxide. After washing in water, the counter was stained by hematoxylin.
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