Kidney, liver, and colon tissues excised from mice were embedded in Richard-Allan Scientific™ Neg-50™ frozen section medium (Thermo Scientific) followed by sectioning at a thickness of 10μm using a Cryostat microtome. For immunohistochemistry (IHC) staining, the slides were treated with 0.1% H2O2, followed by blocking with 10% normal goat serum (NGS). Macrophage staining was performed using a rat anti-mouse F4/80 antibody (BioLegend) at 4°C (16 h) and an HRP-conjugated goat anti-rat secondary antibody, followed by color development using 3, 3′-diaminobenzidine (DAB) (BD Biosciences). Tissue counterstaining was performed using hematoxylin. For immunofluorescence (IF) staining, NGS-blocked tissue sections were incubated with Alexa Fluor 488-conjugated anti-mouse F4/80 antibody (BioLegend) or a non-conjugated anti-F4/80 antibody followed by detection using Alexa Fluor 594-conjugated goat anti-rat IgG (Invitrogen). To test macrophage proliferation in situ, tissue sections were permeabilized with 0.05% Triton X-100, and stained with a FITC-conjugated anti-mouse Ki-67 antibody (BioLegend).
Rat anti mouse f4 80 antibody
Manufactured by BioLegend
The Rat anti-mouse F4/80 antibody is a laboratory reagent used to detect the F4/80 antigen, which is expressed on the surface of mouse macrophages. This antibody can be used in various immunological techniques, such as flow cytometry and immunohistochemistry, to identify and study macrophage populations in mouse tissue samples.
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2 protocols using rat anti mouse f4 80 antibody
1
Immunohistological Analysis of Tissue Macrophages
2
Fluorescence Imaging of Macrophage Activation
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Fixed SVF cells from mice intraperitoneally administered IR-61 for 24 h were incubated with rat anti-mouse F4/80 antibody (#123101, Biolegend; dilution: 1:200) overnight at 4 °C followed by goat anti-rat Alexafluor 488 secondary antibody (#A-11006, Invitrogen; dilution: 1:500). Nuclei were stained using DAPI. Cells were imaged using a confocal fluorescence microscope (Leica TCS SP5). For IR-61 subcellular localization, PMs were incubated with 10 μM IR-61 for 30 min and then stained with Mito-tracker Green (#C1048, Beyotime) for another 30 min. The nuclei were stained by Hoechst33258 for 5 min then imaged by a confocal fluorescence microscope (Leica TCS SP5).
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