Tumor tissues were embedded in Tissue-Tek O.C.T. compound (Sakura Finetek, Nagano, Japan) and frozen. Section slices were placed on glass slides and fixed with cold methanol for 10 min, dried, and then blocked with Block Ace buffer (DS Pharma Biomedical, Osaka, Japan) for 20 min at room temperature. Rabbit anti-mouse CD11b antibody, rat anti-F4/80 antibody (Abcam, Cambridge, UK), and anti-mouse CD31 antibody diluted with Block Ace buffer were added to the slides, which were then incubated for 60 min at room temperature. The slides were then washed three times with PBS. Anti-rat IgG-Alexa488, and anti-rabbit IgG-Alexa594, diluted with Block Ace buffer, were then added to the slides, which were again incubated for 60 min at room temperature before being washed three times with PBS. Image scanning was conducted on a Leica TCS SP8 (Leica Microsystems, Wetzlar, Germany) or NanoZoomer (Hamamatsu Photonics, Shizuoka, Japan); the images were analyzed on a LAS AF (Leica Microsystems) or HALO (Indica Labs, Corrales, NM, USA).
Rat anti f4 80 antibody
Manufactured by Abcam
Sourced in United States
The Rat anti-F4/80 antibody is a laboratory reagent used for the identification and detection of the F4/80 antigen, which is a transmembrane glycoprotein expressed on the surface of mature mouse macrophages. This antibody can be used in various immunological techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to study macrophage biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Tissue tek oct compound, by Sakura Finetek (1 mentions)
Las af, by Leica (1 mentions)
Tcs sp8, by Leica (1 mentions)
Nanozoomer, by Hamamatsu Photonics (1 mentions)
Shandon cytospin 4, by Thermo Fisher Scientific (1 mentions)
Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Goat anti cd206 antibody, by R&D Systems (1 mentions)
Rabbit anti inos antibody, by Abcam (1 mentions)
Enzychrom nadp nadph assay kit, by BioAssay Systems (1 mentions)
Mouse anti gapdh antibody, by Merck Group (1 mentions)
Masson s trichrome staining kit, by Merck Group (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Thermo Fisher Scientific (1 mentions)
Goat anti col1a1, by Santa Cruz Biotechnology (1 mentions)
Anti seh, by Santa Cruz Biotechnology (1 mentions)
Hydroethidine dhe, by Thermo Fisher Scientific (1 mentions)
Anti cd45 fitc, by Thermo Fisher Scientific (1 mentions)
Anti f4 80 pe, by Thermo Fisher Scientific (1 mentions)
Apc anti cd206, by Thermo Fisher Scientific (1 mentions)
Pe cy7 anti cd86, by Thermo Fisher Scientific (1 mentions)
Percp cy5.5 anti cd11b, by Thermo Fisher Scientific (1 mentions)
4 protocols using rat anti f4 80 antibody
1
Immunofluorescent Staining of Frozen Tumor Tissue
2
Immunostaining of Liver Tissue Sections
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DMSCs were attached to slides by centrifugation at 300 r.p.m. for 5 min using Shandon Cytospin 4 (Thermo Fisher, USA). Frozen sections (5 μm) of the liver tissue were used for immunofluorescence staining. Sections were fixed with cold methanol for 10 min and treated with Triton-X 100 for 10 min. Then, they were blocked with donkey serum or goat serum for 30 min at room temperature. Subsequently, sections were incubated with goat anti-CD206 antibody (1 : 200, R&D), rabbit anti-iNOS antibody (1 : 400, Abcam), rat anti-F4/80 antibody (1 : 200, Abcam), rabbit anti-cleaved caspase 3 (1 : 400, Cell Signaling Technology), or rabbit anti-Cathepsin B (1 : 200, Abcam) overnight at 4 °C. Then, sections were incubated with F488-labeled goat anti-rabbit secondary antibody (1 : 400, Thermo Fisher), 555-labeled goat anti-rat secondary antibody (1 : 400, Thermo Fisher), or F488-labeled donkey-anti-goat secondary antibody (1 : 400, Thermo Fisher) for 1 h at room temperature. The results were observed with a fluorescence microscope (Leica, Germany).
3
Antibody Panel for Tissue Analysis
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Rabbit anti-collagen (COL)4A2, anti-sEH, goat anti-COL1A1, mouse anti-α-actin, and rat anti-CD3 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-inducible nitric oxide synthase (iNOS) and anti-myeloperoxidase (MPO) antibodies were from Cell Signaling Technology (Beverly, MA, USA). Rat anti-F4/80 antibody was from Abcam (Cambridge, MA, USA). Mouse anti-GAPDH antibody and Masson's trichrome staining kits were from Sigma-Aldrich (St. Louis, MO, USA). Periodic acid-Schiff staining kit was from Muto Pure Chemical (Tokyo, Japan). Enzyme-linked immunosorbent assay (ELISA) kits for monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), interleukin- (IL-) 1β, and IL-6 were from R&D Systems (Minneapolis, MN, USA). Quantitative assay kit for collagen Sirius Red Staining was from Amsbio (Lake Forrest, CA, USA). Hydroethidine (DHE) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were from Molecular Probes (Eugene, OR, USA). EnzyChrom NADP+/NAD(P)H assay kit was from BioAssay Systems (Hayward, CA, USA).
4
Cardiac Macrophage Phenotyping in MI/R Mice
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In vivo, the randomly divided three groups (n = 6) of MI/R induced mice were treated with 200 µL PBS, LM‐miRs or PLM‐miRs, respectively. Two days after administration (the 5th d post injury), phenotypes of heart macrophages were detected by immunofluorescence assay and flow cytometry as well. All experimental procedures are the same as previously described. Alexa Fluor 647‐anti‐CD206 antibody (141 711, BioLegend, USA), Rat‐anti‐F4/80 antibody (ab6640, Abcam, Japan) and Alexa Fluor 488‐anti‐Rat secondary antibody (ab150165, Abcam, Japan) were used for immunofluorescence staining in this section. For flow cytometry, FITC‐anti‐CD45 (#561 867), PerCP‐Cy5.5‐anti‐CD11b (#45‐0112‐82), PE‐anti‐F4/80 (#565 410), PE‐Cy7‐anti‐CD86 (#25‐0862‐82), APC‐anti‐CD206 (#17‐2062‐82) were all purchased from eBioscience (USA). ELISA were adopted to detect the concentration of cytokines (TNF‐α, IL‐1β, TGF‐β, and IL‐10) in cardiac tissue homogenate after PBS, LM‐miRs or PLM‐miRs treatment according to the manufacturer's instruction. Before the detection, the BCA assay was used to quantify the protein concentration of the tissue homogenate, and the subsequent detection results were also corrected according to the corresponding protein concentration.
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