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F4 80 pe cy5

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F4/80-PE-Cy5 is a fluorescently-labeled antibody that binds to the F4/80 antigen, which is expressed on the surface of mouse macrophages. It can be used for the identification and analysis of mouse macrophage populations in flow cytometry applications.

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Lab products found in correlation

Facscalibur flow cytometer, by BD (2 mentions) Cd11b pe, by BD (2 mentions) Cd45 apc, by BD (2 mentions) Ly6g fitc, by BD (2 mentions) Mouse fc block, by BD (2 mentions) Rbc lysis buffer, by BioLegend (2 mentions) Ly6c apc, by Thermo Fisher Scientific (2 mentions) Fc block, by BD (2 mentions) Foxp3 pe, by Thermo Fisher Scientific (2 mentions) Cd19 pe, by Thermo Fisher Scientific (2 mentions) Cd4 fitc, by Thermo Fisher Scientific (2 mentions) Lsrfortessa, by BD (2 mentions) Cd4 pe cy5, by Thermo Fisher Scientific (2 mentions) Recombinant murine m csf, by R&D Systems (1 mentions) Anti actin antibody, by Merck Group (1 mentions) Tumor necrosis factor (tnf), by R&D Systems (1 mentions) Pe gr1, by Thermo Fisher Scientific (1 mentions) Pecy7 cd11b, by Thermo Fisher Scientific (1 mentions) Rankl, by R&D Systems (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

Close spelling variants of this product

F4/80 (353 citations) F4/80-PE (78 citations) Anti-F4/80 PE-Cy5 (4 citations) F4/80-PE-Cy5 (BM8, (3 citations) PE-Cy5 conjugated anti-F4/80 (2 citations)

11 protocols using f4 80 pe cy5

1

Quantifying Immune Cell Subsets in Pneumococcal Infection

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The MELF of 5 mice from each group at day 1 and day 3 after S.pn inoculation were pooled, centrifuged at 500 g. The cell pellets were treated with RBC lysis buffer (Biolegend, San Diego, United States) and nonspecific binding was blocked with Mouse BD Fc Block in stain buffer (BD Pharmingen, San Jose, United States). A cocktail of fluorophore-conjugated rat anti-mouse cell surface antibodies included APC-CD45 (BD Pharmingen, San Jose, United States), PE-Cy5-F4/80 (eBioscience, San Diego, United States), FITC-Ly6G (BD Pharmingen, San Jose, United States), PE-CD11b (BD Pharmingen, San Jose, United States), and their isotype antibodies were added for 45 min on the ice. After that, the cells were washed three times and resuspended in stain buffer prior to FACS analysis on a BD FACSCalibur flow cytometer (BD Biosciences, San Jose, United States).
Wang Z., He Q., Zhang X., Ma Y., Fan F., Dong Y., Xu W., Yin Y, & He Y. (2018). Innate Anti-microbial and Anti-chemotaxis Properties of Progranulin in an Acute Otitis Media Mouse Model. Frontiers in Immunology, 9, 2952.
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2

Murine Osteoclastogenesis Assay

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Recombinant murine M-CSF, RANKL, TNF were purchased from R&D Systems (Minneapolis, MN). Antibodies of RelB for Western blot were purchased from Santa Cruz and anti-actin antibody was from Sigma. Goat anti-rabbit (or mouse) IgG-HRP conjugate secondary antibody was purchased from Bio-Rad. Fluorescent-labeled rat anti-mouse antibodies APC-Ly6C, PEcy7-CD11b, PE-Gr1 (also called Ly6G) and PECy5-F4/80 were purchased from eBioscience. Ammonium Chloride (NH4Cl) solution was purchased from STEMCELL technologies.
Zhao Z., Hou X., Yin X., Li Y., Duan R., Boyce B.F, & Yao Z. (2015). TNF Induction of NF-κB RelB Enhances RANKL-Induced Osteoclastogenesis by Promoting Inflammatory Macrophage Differentiation but also Limits It through Suppression of NFATc1 Expression. PLoS ONE, 10(8), e0135728.
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3

Immune Cell Profiling of Spn-Infected Mice

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The MELF of five mice from each group at 1 and 3 d after Spn inoculation were pooled, centrifuged at 500 g. The cell pellets were treated with RBC lysis buffer (Biolegend, San Diego, CA) and nonspecific binding was blocked with Mouse BD Fc Block in stain buffer (BD Pharmingen, Franklin Lakes, NJ). A cocktail of fluorophore-conjugated rat anti-mouse cell surface antibodies included APC-CD45 (BD Pharmingen), PE-Cy5-F4/80 (eBioscience, San Diego, CA), FITC-Ly6G (BD Pharmingen), PE-CD11b (BD Pharmingen), and their isotype antibodies were added for 45 min on the ice. After that, the cells were washed three times and resuspended in stain buffer prior to FACS analysis on a BD FACS Calibur flow cytometer (BD Biosciences, San Jose, CA).
Huang Y., Wang Z., Jin C., Wang L., Zhang X., Xu W., Xiang Y., Wang W., He X., Yin Y, & He Y. (2016). TLR2 promotes macrophage recruitment and Streptococcus pneumoniae clearance during mouse otitis media. Pediatric research, 80(6).
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4

Multicolor Flow Cytometry for Immune Cell Profiling

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Cells (3×105–5×105/well) were fist incubated in FcBlock (BD Pharmingen) and then with cocktails of antibodies using CD45-APCCy7 (BD Pharmingen, 30-F11), F4/80-PECy5 (eBioscience, BM8), CD11b-AlexaFluor700 (BD Pharmingen, M1/70), MHCII-biotin (eBiosciences, M5/114.15.2) followed by streptavidin-PETexasRed (BD Pharmingen), CD64-PE (BD Pharmingen, X54-5/7.1.1), CX3CR1-unconjugated (AbD Serotec, polyclonal) followed by anti-IgG(H+L)-FITC (Southern Biotech), and anti-IL10-APC (eBiosciences, JES5-16E3), CD3-PECy5 (eBiosciences, 145-2C11), CD4-PECy7 (eBiosciences, GK1.5), CD19-PE (eBiosciences, MB19-1), FoxP3-PE (eBiosciences, FJK-16s), and PD1-PE (eBiosciences, J43). For intracellular staining, cells were fixed and permeabilized with Cytofix-Cytoperm solution (eBiosciences). Flow results were computed with a BD LSR II flow cytometer and data analyses was performed by using the FACS Diva software (BD).
Viladomiu M., Bassaganya-Riera J., Tubau-Juni N., Kronsteiner B., Leber A., Philipson C.W., Zoccoli-Rodriguez V, & Hontecillas R. (2017). Cooperation of gastric mononuclear phagocytes with Helicobacter pylori during colonization. Journal of immunology (Baltimore, Md. : 1950), 198(8), 3195-3204.
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5

Isolation and Identification of Peritoneal Macrophages

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Freshly isolated PEC were washed and resuspended in a volume of 1 ml. The cells were incubated with Fc block (BD Biosciences; 5 μl/1 ml) for 10 min to prevent non-specific binding. For F4/80+ CD11b+ macrophages, PEC were surface stained with 1 μl per 5 × 106 cells of fluorescent antibodies directed against CD11b-APC-eFluor780 and F4/80-PECy5 (eBioscience, UK) for 15 min at room temperature (RT) in the dark. For small peritoneal macrophages (SPM), PEC were surface stained with 1 μl per 5 × 106 cells of fluorescent antibodies directed against CD11b, F4/80 and MHC II-FITC (eBioscience, UK) for 15 min at RT in the dark. Cells were washed and resuspended in 1 ml of FACS buffer and sorted by flow cytometry. SPM were identified as CD11b+ F4/80low MHCIIhigh cells.
Quinn S.M., Cunningham K., Raverdeau M., Walsh R.J., Curham L., Malara A, & Mills K.H. (2019). Anti-inflammatory Trained Immunity Mediated by Helminth Products Attenuates the Induction of T Cell-Mediated Autoimmune Disease. Frontiers in Immunology, 10, 1109.
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6

Spleen Cell Profiling by Flow Cytometry

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Spleens were harvested at day 3 PI, and single cell suspensions were prepared and fixed in 4% PFA. Non-specific binding was blocked using a rat anti-mouse antibody against the FcγIII/II receptor (CD16/CD32) (BD), and the following rat anti-mouse cell surface antibodies were used: B220-PE-Cy7 (BD), CD4-FITC (eBioscience), CD8a-PE-Cy5 (BD), CD11c-APC (eBioscience), CD11b-PE-Cy7 (eBioscience), Ly6G-PE (BD), and F4/80-PE-Cy5 (eBioscience). For intracellular staining, the following antibodies were applied: rat anti-mouse CD68-Alexa700 (BioRad), rabbit anti-mouse iNOS (AbCam) with goat anti-rabbit-Alexa488 (Invitrogen). Fluorescent signals were detected using a LSRii flow cytometer (BD). See also Supplemental Experimental Procedures.
Davis K.M., Mohammadi S, & Isberg R.R. (2014). Community behavior and spatial regulation within a bacterial microcolony in deep tissue sites serves to protect against host attack. Cell host & microbe, 17(1), 21-31.
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7

Immune Cell Analysis Protocol

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The immune cells were analyzed following the standard protocol. Briefly, tumors were collected and incubated in dissociation buffer with 1640 medium (contained collagenase, hyaluronidase, and deoxyribonuclease I) at 37 °C for digest tumor tissues to the single‐cell suspension. And then the cells were stained with surface antibodies: CD3‐PE (eBioscience, Catalog: CD0304), CD4‐FITC (eBioscience, Catalog: 11‐0041‐82), CD8‐PerCP‐Cy5.5 (eBioscience, Catalog: 45‐0081‐82), CD86‐PE (eBioscience, Catalog: 12‐0862‐81), CD11b‐FITC (eBioscience, Catalog: 11‐0112‐81), F4/80‐PE‐Cy5 (eBioscience, Catalog: 15‐4801‐82), CD206‐PE (eBioscience, Catalog: 12‐2061‐82), CD44‐PE (eBioscience, Catalog: 12‐0441‐81), or CD62L‐APC (eBioscience, Catalog: 17‐0621‐81) respectively for 30 min. After fixing and perforating the cells, the intracellular markers: FOXP3‐PE (eBioscience, Catalog: 12‐5773‐82) or IFN‐γ‐PE (eBioscience, Catalog: 12‐7311‐82) were stained. The stained cells were detected using flow cytometry (Beckman Cytoflex S flow cytometer). The data were analyzed using FlowJo 10.0.
Sang W., Xie L., Wang G., Li J., Zhang Z., Li B., Guo S., Deng C, & Dai Y. (2020). Oxygen‐Enriched Metal‐Phenolic X‐Ray Nanoprocessor for Cancer Radio‐Radiodynamic Therapy in Combination with Checkpoint Blockade Immunotherapy. Advanced Science, 8(4), 2003338.
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8

Flow Cytometry Analysis of Mouse Tumor Tissue

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Small samples of mouse tumor tissues (2–3 wk after last immunization) were chopped in 1 ml of cell dissociation solution (1% 100 mg/ml collagenase/dispase, 0.5% 20 mg/ml DNase in DMEM) and incubated for 30 min at 37°C. The cell suspension (1×105 to 1×106 cells) was stained for use in flow cytometry after blocking the Fc receptors by applying ultra-blocking buffer (10% normal mouse serum, 10% normal hamster serum, 10% normal rat serum, and monoclonal Ab 2.4G2 in PBS). Abs used were CD11c-FITC (eBioscience, San Diego, CA, USA), CD45-PE-Texas-Red (Invitrogen, Carlsbad, CA, USA), F4/80-PE-Cy5 (eBioscience), CD11b-biotin (Miltenyi, Bergisch Gladbach, Germany), Gr-1-PE-Cy7 (eBioscience), and SA-PerCP-Cy5.5 (eBioscience). All samples were filtered through a 200 μm nylon mesh before running in BD FACSAria. The results were analyzed by using the Flowjo 10 program (Tree Star, Inc., Ashland, OR, USA).
Kim K.Y., Cho E.H., Yoon M, & Kim M.G. (2020). Critical Adjuvant Influences on Preventive Anti-Metastasis Vaccine Using a Structural Epitope Derived from Membrane Type Protease PRSS14. Immune Network, 20(4), e33.
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9

Quantifying Macrophage Surface Markers

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Peritoneal macrophages were harvested from WT and β3−/− mice as described in Animal Models. For adenoviral receptors, cells were blocked with Fc fragments for 15min before addition of conjugated antibodies (CAR-PE (Millipore, Watford, UK), β3-PE (eBioscience, Hatfield, UK), β5-PE (eBioscience, Hatfield, UK) and IgG-PE (eBioscience, Hatfield, UK)) in PBS+1% BSA for 1hour on ice in the dark. Samples were analysed on a BD FACSCalibur (BectonDickinson, Oxford, UK).
To quantify macrophage cell surface markers using multichannel flow cytometry, cells were incubated with a viability dye (eBioscience, Hatfield, UK) for 30min in the dark on ice and then blocked with Fc fragments for 15min. Conjugated antibodies (CD45-FITC (eBioscience, Hatfield, UK), CD11b-eFluor450 (eBioscience, Hatfield, UK), mMR-PE/CD206 (BioLegend, London, UK), Ly-6C-APC (eBioscience, Hatfield, UK), Ly-6G-AlexaFluor700 (eBioscience, Hatfield, UK), CsfR1/CD115 (eBioscience, Hatfield, UK) and F4/80-PE-Cy5 (eBioscience, Hatfield, UK)) were added for 15min on ice in the dark. Cells were washed and fixed in 2% formaldehyde. Samples were analysed on a BD LSRFortessa (BectonDickinson, Oxford, UK).
Browne A., Tookman L.A., Ingemarsdotter C.K., Bouwman R.D., Pirlo K., Wang Y., McNeish I.A, & Lockley M. (2015). Pharmacological inhibition of β3 integrin reduces the inflammatory toxicities caused by oncolytic adenovirus without compromising anticancer activity. Cancer research, 75(14), 2811-2821.
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10

Multicolor FACS Analysis of Immune Cells

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FACS analyses for different immune cell populations were conducted following standard protocols. Freshly-harvested splenocytes and peripheral blood mononuclear cells (PBMCs) were stained with the immune cell markers listed below and analyzed with BD FACS Diva on LSR II. Antibodies used for FACS analysis: CD3 Alexa700 (eBioscience), CD4 PE-Cy5 (eBioscience), CD8 Alexa488 (eBioscience), CD44 APC (eBioscience), CD62L PE-Cy7 (eBioscience), CD19 PE (eBioscience), CD11b Pe-eFluor 610 (eBioscience), F4/80 PE-Cy5 (eBioscience), Ly-6G(ar-1) APC (eBioscience), CD45 APC-eFluor 780 (eBioscience).
Rangan P., Choi I., Wei M., Navarrete G., Guen E., Brandhorst S., Enyati N., Pasia G., Maesincee D., Ocon V., Abdulridha M, & Longo V.D. (2019). Fasting-Mimicking Diet Modulates Microbiota and Promotes Intestinal Regeneration to Reduce Inflammatory Bowel Disease Pathology. Cell reports, 26(10), 2704-2719.e6.
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