Cells collected from BALF were incubated in 100 μl 1×RBC lysis buffer (BioLegend) on ice for 10 min, and then quenched by adding 1 ml PBS. Cells from each sample were resuspended in 100 μl Stain Buffer (554656, BD Pharmingen) and incubated for 15 min on ice with 1 μg mouse IgG (ab37355, Abcam, 1:500) for Fc blocking. Then cells were incubated with primary monoclonal antibodies including rat anti-mouse Gr1/Ly-6G-Alexa Fluor 594 (Novus, 1:200), rat anti-mouse CD11b-FITC (R&D System,1:200) and rat anti-F4/80-APC (R&D System,1:200). Flow cytometry was performed using FACS LSR Fortessa X20 (BD Biosciences) and data were analyzed by BD FACSDiva 8.0.1 and FlowJo v10.8.1.
Rat anti f4 80 apc
Manufactured by R&D Systems
Rat anti-F4/80-APC is a fluorochrome-conjugated antibody directed against the F4/80 antigen, a glycoprotein expressed on the surface of mouse macrophages and microglia. This antibody can be used to identify and quantify macrophage populations in flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti mouse gr1 ly 6g alexa fluor 594, by Novus Biologicals (2 mentions)
Rat anti mouse cd11b fitc, by R&D Systems (2 mentions)
Stain buffer, by BD (1 mentions)
Rbc lysis buffer, by BioLegend (1 mentions)
Facs lsr fortessa x20, by BD (1 mentions)
Ab37355, by Abcam (1 mentions)
Lsm 710 confocal microscope, by Zeiss (1 mentions)
Ht5014, by Merck Group (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
2 protocols using rat anti f4 80 apc
1
Immune Cell Profiling in BALF
2
Immunofluorescent Staining of Mouse Lung Tissue
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Lung lobes were fixed with formalin solution (HT5014, Sigma) at room temperature for 5 min. Then the slides were blocked for 1 h at room temperature by 5% horse serum in 0.3% TritonX-100/PBS and then incubated overnight at 4 °C with the appropriate primary antibodies. The nuclei were stained with ProLong™ Gold Antifade Mountant with DAPI (ThermoFisher). The antibodies used for IF staining includes rat anti-mouse Gr1/Ly-6G-Alexa Fluor 594 (Novus, 1:200), rat anti-mouse CD11b-FITC (R&D System,1:200) and rat anti-F4/80-APC (R&D System,1:200). The sections were washed with PBS, dried at room temperature and observed via LSM710 confocal microscope (Zeiss).
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