Primary rat antimouse f4 80 antibody

Manufactured by Bio-Rad

Sourced in United States

The Primary rat antimouse F4/80 antibody is a laboratory tool used for the identification and isolation of mouse F4/80-positive cells, which are often macrophages. The antibody recognizes the F4/80 cell surface antigen expressed on various mouse myeloid cell types.

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Lab products found in correlation

Ar9 buffer, by Akoya Biosciences (1 mentions) Rat anti mouse f4 80, by Bio-Rad (1 mentions) Opal 520 reagent pack, by PerkinElmer (1 mentions) Opal 570 reagent pack, by PerkinElmer (1 mentions) Rabbit antimouse ym1, by STEMCELL (1 mentions) Blocking diluent buffer, by PerkinElmer (1 mentions) Eclipse 90i, by Nikon (1 mentions) Primary rabbit anti mouse α sma antibody, by Abcam (1 mentions) Goat anti rabbit igg hrp secondary antibody, by Abcam (1 mentions) Bond polymer refine detection system, by Leica (1 mentions)

2 protocols using primary rat antimouse f4 80 antibody

Immunohistochemical Analysis of Murine Macrophages

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Formalin-fixed, paraffin-embedded tissue sections (pancreatic body and tail; female and male mice combined) were dewaxed with xylene and rehydrated in an ethanol series. Antigen retrieval was performed in AR9 buffer (Akoya Biosciences, Menlo Park, CA), and sections were then immersed in blocking/diluent buffer (PerkinElmer, Richmond, CA). For immunohistochemistry, sections were incubated overnight with primary rat antimouse F4/80 antibody (#MCA497B; Bio-Rad) diluted 1:200 in blocking/diluent buffer. After washing, the Labelled Polymer-Dako REAL EnVision HRP kit (Agilent Dako, Santa Clara, CA) was then used to visualize antigen-antibody complexes.
For immunofluorescence, sections were incubated overnight with rat antimouse F4/80 (#MCA497B; Bio-Rad), rabbit antimouse pSTAT (Tyr701) (#44-376; Thermo Fisher Scientific), and rabbit antimouse Ym1 (#60130; STEMCELL Technologies, Cambridge, MA) antibodies (1:200 dilution in blocking buffer). Slides were then counterstained with multiplexing fluorescence kits from PerkinElmer: Opal 520 Reagent Pack, FITC, Opal 570 Reagent Pack, TRITC, and Opal 650 Reagent Pack, CY5, according to manufacturer’s instructions. 4′,6-diamidino-2-phenylindole was used for nuclear counterstain. Images were taken with a Nikon Eclipse 90i.

Ako S., Teper Y., Ye L., Sinnett-Smith J., Hines O.J., Rozengurt E, & Eibl G. (2022). Statins Inhibit Inflammatory Cytokine Production by Macrophages and Acinar-to-Ductal Metaplasia of Pancreatic Cells. Gastro hep advances, 1(4), 640-651.

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Immunohistochemical Analysis of Macrophages and Myofibroblasts

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Paraffin‐embedded tissue sections were deparaffinized, rehydrated and pretreated with sodium citrate (0.05%) to unmask the antigen, followed by incubation with H₂O₂ (0.3%) for 25 min at room temperature in dark conditions, to block endogenous peroxidase activity, and with 2% BSA for 20 min at room temperature, to avoid nonspecific binding of the primary antibody. Thereafter, the liver sections were incubated 1 h at room temperature with a primary rat anti‐mouse F4/80 antibody (dilution 1:100; Bio‐Rad, Hercules, CA, USA) followed by incubation for 15 min at room temperature with a biotinylated rabbit anti‐rat IgG secondary antibody (Abcam) and 30 min with HRP goat anti‐rabbit IgG secondary antibody (dilution 1:200; Abcam). For α‐smooth muscle actin (α‐SMA), sections were incubated with primary rabbit anti‐mouse α‐SMA antibody (dilution 1:250; Abcam) for 1 h with HRP goat anti‐rabbit IgG secondary antibody (dilution 1:200; Abcam) for 30 min. The signal was detected by the Bond Polymer Refine Detection system (Leica Biosystems, Wetzlar, Germany). The sections were visualized at 200×, and positive staining for F4/80 and α‐SMA was quantified by histomorphometry. The results are expressed as a percentage of the F4/80‐positive or α‐SMA‐positive area.

Flores‐Costa R., Alcaraz‐Quiles J., Titos E., López‐Vicario C., Casulleras M., Duran‐Güell M., Rius B., Diaz A., Hall K., Shea C., Sarno R., Currie M., Masferrer J.L, & Clària J. (2018). The soluble guanylate cyclase stimulator IW‐1973 prevents inflammation and fibrosis in experimental non‐alcoholic steatohepatitis. British Journal of Pharmacology, 175(6), 953-967.

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