F4 80 pacific blue

Manufactured by BioLegend

Sourced in Germany

The F4/80-Pacific Blue is a fluorescence-labeled antibody that binds to the F4/80 antigen, a cell surface glycoprotein expressed primarily on mature mouse macrophages. This product can be used for the identification and enumeration of macrophages in flow cytometry applications.

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Lab products found in correlation

4 protocols using f4 80 pacific blue

Multiparametric Flow Cytometry Profiling of Hematopoietic Lineages

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Blood lymphoid. CD4-Fitc (BioLegend), CD8-PE-Cy7 (BD), B220–Pacific blue (BD), CD11b-PerCpCy5,5 (BD), CD3-APC (BioLegend); Blood myeloid and erythroid: CD3-Fitc (BD), CD19-Fitc (BD), B220-Fitc (BD), Gr1-PE (BD), F4/80-Pacific blue (BioLegend), CD11b-APC (BD);
BM lymphoid. IgD-Fitc (BD), CD25-PE (BioLegend), CD11b-PerCpCy5,5 (BD), IgM-PECy7 (Southern Biotech), B220-Pacific blue (BD), cKit-APC (BD); BM myeloid: Gr1-PE (BD), CD11b-PerCpCy5,5 (BD), Ter119-PECy7 (BD), F4/80–Pacific blue (BioLegend), CD19-APC (BioLegend); BM megakaryocyte: CD3-Fitc (BD), CD41-PE (BioLegend), CD11b-PerCpCy5,5 (BD), B220–Pacific blue (BD), CD19-APC (BioLegend); HSPC and CLP: Strep-APC/CY7 (Southern Biotech), cKit-APC (BD), SCA1-PacBlue (BioLegend), CD34-FITC (eBioscience), CD135-PE (BioLegend), CD150-PECy7 (BioLegend), CD127-PErCpCy5,5 (BioLegend); Hematopoietic progenitors: Strep-APC/CY7 (Southern Biotech), cKit-APC (BD), SCA1-PacBlue (BioLegend), CD34-FITC (eBioscience), CD16/32-PerCpCy5,5 (eBioscience), CD127-PE (eBioscience).

Alendar A., Lambooij J.P., Bhaskaran R., Lancini C., Song J.Y., van Vugt H., Snoek M, & Berns A. (2020). Gene expression regulation by the Chromodomain helicase DNA-binding protein 9 (CHD9) chromatin remodeler is dispensable for murine development. PLoS ONE, 15(5), e0233394.

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Flow Cytometry Analysis of Tumor-Infiltrating Lymphocytes

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Flow cytometry analysis was performed by using a BD FACSVerse (BD Biosciences, Milan, Italy) and the following monoclonal anti-human antibodies: CD45-APC-H7, CD3ɛ-PerCP, CD3ɛ-FITC, CD8α-V500, CD56-V450, CD56-PE-Cy7, CD19-FITC, IFN-γ-FITC, IFN-γ-PE, IL-17A-V450, IL-17F-PE, TNFα-PE (all from BD Pharmingen, Milan, Italy), IL-21-PE, IL-22-APC, IL-6-PerCP, T-bet-PE-Cy7 (clone 4B10), RORγt-APC (clone AFKJS-9), RORγt-PE (clone AFKJS-9) (all from eBioscience) and CD68 (BioLegend, San Diego, CA, USA). TILs isolated from mouse colon tumors were stained with the following antibodies: CD3ɛ-Pacific Blue, CD8α-FITC, CD45-APC-Cy7, CD49b-PE (clone DX5), CD19-APC (all from BD Pharmingen) and F4/80-Pacific Blue (BioLegend). In parallel, cells were stained with the respective control isotype antibodies.

De Simone V., Franzè E., Ronchetti G., Colantoni A., Fantini M.C., Di Fusco D., Sica G.S., Sileri P., MacDonald T.T., Pallone F., Monteleone G, & Stolfi C. (2014). Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth. Oncogene, 34(27), 3493-3503.

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Multiparameter Flow Cytometry Immune Profiling

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Nonspecific antibody binding was blocked with FcR Blocking Reagent (Miltenyi Biotec). Antibodies used for cell-surface staining were CD45-APC-Cy7, F4/80-Pacific Blue (BioLegend, Fell, Germany), Gr-1-APC, CD11b-PerCPVio700, Siglec-f-PE, MHC-Class II-FITC, CD11c-FITC and CD49b-PE (Miltenyi Biotec). Cells were blocked, stained with antibodies according to the manufacturer's protocol, and fixed for 20 min in 2.25% formaldehyde [75] (link). Cells were washed with autoMACS Buffer and analysed using the MACSQuant Analyzer (Miltenyi Biotec). Gating strategy for identification of different subsets was based on Hackstein et al. [77] (link). Briefly, cells were gated based on scatter light (FSC, SSC) characteristics and viable singlet leukocytes were identified by CD45 expression. Out of the CD45-positive cells, neutrophils were identified by Gr-1^bright CD11b^bright expression. Out of the neutrophil negative fraction, eosinophils were identified by Siglec-f⁺ MHC-Class-II^neg expression. Out of the neutrophil and eosinophil negative fraction macrophages were identified as Siglec-f⁺⁺ F4/80⁺ double positive cells. Total dendritic cells were identified as CD11c⁺ Siglec-f^neg CD49b^neg leukocytes. Cell numbers of subsets were calculated per organ.

Bast A., Krause K., Schmidt I.H., Pudla M., Brakopp S., Hopf V., Breitbach K, & Steinmetz I. (2014). Caspase-1-Dependent and -Independent Cell Death Pathways in Burkholderia pseudomallei Infection of Macrophages. PLoS Pathogens, 10(3), e1003986.

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Multiparametric Flow Cytometric Analysis of Intestinal Leukocytes

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Isolated intestinal LP cells were pre-incubated with anti-CD16/CD32 antibodies (Biolegend, San Diego, CA) for 15 min in the dark on ice to prevent non-specific binding of the fluorochrome-conjugated antibodies. The cells were then incubated with the following fluorochrome-conjugated antibodies for 30 min on ice in the dark: F7/4-FITC (1:200; Abcam, Cambridge, MA), Ly-6G (Gr-1)-PECy5 (1:100; Biolegend, San Diego, CA), CD11b-eFluor605 (1:100; eBioscience, San Diego, CA), F4/80-Pacific Blue (1:100; Biolegend, San Diego, CA), CD45-APC-eFluor780 (1:100; eBioscience), Siglec-F PE (1:200; BD Biosciences, Franklin Lakes, NJ), CD19 PerCPCy5.5 (1:200; eBioscience), NK1.1-PECy7 (1:200; Biolegend), CD3-v500 (1:200; BD Biosciences, Franklin Lakes, NJ), and MHCIIc-ef650 (1:200; eBioscience). Dead cells were excluded by propidium iodide (PI) (Sigma-Aldrich, St. Louis, MO) uptake and gated on singlets. Fluorescence minus one (FMO) controls were used for gating. Data were acquired on an LSR II (BD, Franklin Lakes, NJ) and analyzed using FlowJo software (TreeStar Inc., Ashland, OR). A total of n = 7–9 mice/genotype were used for each assay.

Robinson S.C., Chaudhary R., Jiménez-Saiz R., Rayner L.G., Bayer L., Jordana M, & Daniel J.M. (2019). Kaiso-induced intestinal inflammation is preceded by diminished E-cadherin expression and intestinal integrity. PLoS ONE, 14(6), e0217220.

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