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F4 80 apc

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F4/80-APC is a monoclonal antibody that binds to the F4/80 antigen, a glycoprotein expressed on the surface of murine macrophages. It is conjugated with the fluorescent dye Allophycocyanin (APC), enabling the detection and analysis of macrophages in flow cytometry applications.

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Lab products found in correlation

Cd11b fitc, by Thermo Fisher Scientific (8 mentions) Fc block, by BD (7 mentions) Cd4 fitc, by Thermo Fisher Scientific (7 mentions) Facscanto 2, by BD (7 mentions) Cd11b pe, by Thermo Fisher Scientific (6 mentions) Cd11b pe cy7, by Thermo Fisher Scientific (5 mentions) Cd3 pe cy7, by Thermo Fisher Scientific (5 mentions) Cd11c pe cy7, by Thermo Fisher Scientific (5 mentions) Ly6g pe cy7, by BD (4 mentions) Cd8 apc, by Thermo Fisher Scientific (4 mentions) Facscalibur, by BD (4 mentions) Cd11b pe cy7, by BioLegend (4 mentions) Cd11c pe, by BD (4 mentions) Collagenase, by Merck Group (4 mentions) Cd45 fitc, by Thermo Fisher Scientific (4 mentions) Fixable viability dye efluor 506, by Thermo Fisher Scientific (4 mentions) Dnase 1, by Merck Group (4 mentions) Cd4 apc, by Thermo Fisher Scientific (3 mentions) Facscanto 2 flow cytometer, by BD (3 mentions) Cd11c apc, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

F4/80 (353 citations) Anti-F4/80-APC (35 citations) Anti-mouse F4/80-APC (14 citations) F4/80 APC-eFluor 780 (7 citations) APC-conjugated anti-F4/80 (7 citations) Anti-F4/80-APC (clone BM8, (6 citations) APC-conjugated anti-F4/80 (clone BM8, (6 citations) APC-conjugated anti-F4/80 antibody (6 citations) F4/80-APC (clone BM8, (5 citations) APC conjugated anti-mouse F4/80 (clone BM8, (5 citations)

72 protocols using f4 80 apc

1

Multiparameter Flow Cytometry Analysis

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Cell suspensions were treated with anti CD16/CD32 (2.4G2) and then surface stained with combinations of the following fluorochrome-conjugated antibodies: From Biolegend: CD3e–FITC (clone: 145 2C11), Ly6C–FITC or APC/Cy7 (clone: HK1.4), Ly6G–PE/Cy7 (clone: 1A8), CD3e–APC/Cy7 (clone: 145 2C11), CD45R/B220–Pacific Blue (clone: RA3 6B2), CCR2-AlexaFluor647 (clone: SA203G11), CX3CR1-APC (clone: SA011F11), IgG2A,κ-APC isotype control; from BD Biosciences: NK1.1–PerCP/Cy5.5 (clone: PK136); from Ebiosciences: F4/80–APC (clone: BM8), CD4 (L3T4)–FITC (clone: RM4 5), CD11b–FITC (clone: M1/70), CD11b–PE (clone: M1/70), CD4–APC (clone: GK1.5). Cells were acquired on FACSCanto or AriaIII flow cytometers using FACSDiVa software, and data were analyzed using FlowJo software (Tree Star).
Seregin S.S., Golovchenko N., Schaf B., Chen J., Eaton K.A, & Chen G.Y. (2016). NLRP6 function in inflammatory monocytes reduces susceptibility to chemically-induced intestinal injury. Mucosal immunology, 10(2), 434-445.
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2

Phenotyping Extracellular Vesicles by Flow

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Cellular source of EVs was determined by flow cytometry using CD9 Exo-Flow capture beads (System Biosciences; Palo Alto, CA, USA) as per manufacturers' instructions. EVs were attached to CD9 labeled Exo-Flow beads at 4°C, for 16 h, and stained with F4/80-APC, CD31-FITC, CD326-PE, and Ly6G-BV421 (eBioscience) antibodies, and analyzed on BD LSR Fortessa Flow Cytometer at OSU Flow cytometry core facility. Data were analyzed using FlowJo.
Nirujogi T.S., Kotha S.R., Chung S., Reader B.F., Yenigalla A., Zhang L., Shapiro J.P., Wisler J., Christman J.W., Maddipati K., Parinandi N.L, & Karpurapu M. (2022). Lipidomic Profiling of Bronchoalveolar Lavage Fluid Extracellular Vesicles Indicates Their Involvement in Lipopolysaccharide-Induced Acute Lung Injury. Journal of Innate Immunity, 14(5), 555-568.
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3

Tumor Immune Cell Isolation and Characterization

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The tumors were weighed, minced into small fragments, and digested at 37°C in 10 ml of digestion solution [PBS supplemented with type I Collagenase (200 U/ml), Hyaluronidase, and DNase I (100 μg/ml)] for 60 min. Single-cell suspensions were obtained by grinding the digested tissues and filtering them through a 70-μm cell strainer (BD Biosciences). The immune cells were isolated using Ficoll density gradient centrifugation. Freshly isolated immune cells were stained with antibodies for 30 min at 4°C. The following monoclonal anti-mouse antibodies were used: CD45-PECy5.5 (eBioscience), CD3-pecy7 (eBioscience), CD8-APC (eBioscience), NK1.1-APC (eBioscience), F4/80-APC (eBioscience), CD11b-pecy7 (eBioscience), CD11c-APC (eBioscience), and MHCII-PE (eBioscience). Flow cytometry was performed on a FACS Canto II flow cytometer (BD Biosciences), and the data were analyzed using FlowJo software (TreeStar, Ashland, OR).
Xi Q., Chen Y., Yang G.Z., Zhang J.Y., Zhang L.J., Guo X.D., Zhao J.Y., Xue Z.Y., Li Y, & Zhang R. (2020). miR-128 Regulates Tumor Cell CD47 Expression and Promotes Anti-tumor Immunity in Pancreatic Cancer. Frontiers in Immunology, 11, 890.
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4

Macrophage Surface Marker and Intracellular Cytokine Analysis

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For analysis of cell surface markers, the cells were stained with anti-CD86-FITC or anti-CD86-PE, anti-MHC class II-PE-Cy5 (all from BD Biosciences), and F4/80-FITC (Biolegend) or F4/80-APC (eBioscience). Transfection plasmids and siRNA had fluorescent reporters (GFP for plasmid and Cy3-labeled RNA for siRNA), which were used to gate on transfected cells. Macrophages were analyzed for expression of surface markers (MHC class II, CD86, F4/80) or intracellular markers (Egr2, IL-6, TNF). FcRs were blocked with mAb specific for mouse FcR (2.4G2; BD Biosciences). Intracellular staining for Egr2, IL-6 or TNF was performed similarly as described earlier (32 (link), 33 (link)) using fixation/permeabilization agent (eBioscience) and anti-Egr2-APC (eBioscience), anti-IL-6-PE, or anti-TNF-PE mAbs (both from BD Biosciences). The cells were analyzed using LSRFortressaTM cytometer (BD Biosciences) and FlowJo software (Tree Star Inc.) as we described earlier (34 (link)).
Veremeyko T., Yung A.W., Anthony D.C., Strekalova T, & Ponomarev E.D. (2018). Early Growth Response Gene-2 Is Essential for M1 and M2 Macrophage Activation and Plasticity by Modulation of the Transcription Factor CEBPβ. Frontiers in Immunology, 9, 2515.
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5

Flow Cytometry Analysis of Immune Cell Subsets

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Skin samples were prepared as described previously [15 (link)], and flow cytometry was performed as described previously [16 (link)]. In brief, single-cell suspensions (1 million cells) were first incubated with Fc Block (BD Pharmingen, San Diego, CA) for 10 minutes, then coincubated with antibodies for 20 minutes at 4 °C, followed by washing with staining buffer (PBS + 1% FBS). Flow cytometry analysis was performed with an LSRFortessa cell analyzer (BD Biosciences, San Jose. CA), and data were analyzed using Flowjo software (Flowjo, Ashland, OR). CD19-FITC (BD Pharmingen), CD229.1-PE (BD Pharmingen), CD4-PE-Cy7 (eBioscience, San Diego, CA), CD25-APC-Cy7 (BD Biosciences), CD45-PB (BioLegend San Diego, CA), CD8α-AF700 (BioLegend), B220-PE (BD Biosciences), CD138-PE (BioLegend). CD11c-FITC (BioLegend). F4/80-APC (eBioscience), and CD11 b-APC-Cy7 (BioLegend) were used for these studies.
Pai C.C., Khuat L.T., Chen M., Murphy W.J, & Abedi M. (2016). Therapeutic Effects of a NEDD8-Activating Enzyme Inhibitor, Pevonedistat, on Sclerodermatous Graft-versus-Host Disease in Mice. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, 23(1), 30-37.
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6

Multiparametric Analysis of Liver-Derived Immune Cells

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A nonparenchymal cell fraction from whole liver extracts was isolated upon collagenase and mechanical digestion followed by Percoll (GE Healthcare Life Sciences) gradient centrifugation as previously described [5 (link)]. In parallel, blood samples were collected in EDTA-containing tubes and treated with red blood cell lysis buffer (PharmLyse, BD Biosciences, Germany). Upon removal of red bodies and centrifugation, immune cells were incubated with fluorochrome-conjugated antibodies and characterized according to two different panels, a myeloid panel: CD45-BV510 (103138, BioLegend), 7AAD-PE-Cy5-YG (420404, BioLegend), CD11b-BV711 (101242, BioLegend), F4/80-APC (17-4801-82, eBiosciences), MHC2-Alexa700 (107622, BioLegend), CD11c-PE-Cy7 (25-0114-81, eBiosciences), and Ly6G-FITC (551460, BD Pharmingen) and a lymphoid panel: CD45-BV510 (103138, BioLegend), 7AAD-PE-Cy5-YG (420404, BioLegend), CD3-PE-Cy7 (25-0031-82, eBiosciences), CD4-FITC (11-0041-85, eBiosciences), CD8-PerCpCy5.5 (126610, BioLegend), and NK1.1-BV711 (108745, BioLegend). Labeled cells were then subjected to flow cytometry using a BD Canto II (BD Biosciences) and relative cell numbers were analyzed using FlowJo software (Tree Star).
Ramadori P., Drescher H., Erschfeld S., Fragoulis A., Kensler T.W., Wruck C.J., Cubero F.J., Trautwein C., Streetz K.L, & Kroy D.C. (2017). Genetic Nrf2 Overactivation Inhibits the Deleterious Effects Induced by Hepatocyte-Specific c-met Deletion during the Progression of NASH. Oxidative Medicine and Cellular Longevity, 2017, 3420286.
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7

Flow Cytometry Analysis of Immune Cells

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Stained cells were analyzed using a BD LSR II-SORP system (Becton Dickinson) and analyzed with FlowJo software (Tree Star). The following mAbs were used: CD45-PerCPCy5, CD45-APC, CD8-APC, anti-IFNγ-PECy7 and anti-IL-4-FITC (BD Biosciences), Gr1-APC, CD11c-PECy7, CD11b-PB, F4/80-APC (eBiosciences), 1A8-APCCy7, Ly6C-FITC, 1A8-APCCy7, CD4-AF700. DAPI and AnV-FITC (BioLegends). In vivo bioluminescence imaging of MPO activity was quantified through injection of 200mg/kg luminol (Carbosynth) i.p. 10 minutes before luminescence acquisition. Photon emission was acquired for 10 minutes using a Xenogen IVIS Imaging system.
Hurrell B.P., Schuster S., Grün E., Coutaz M., Williams R.A., Held W., Malissen B., Malissen M., Yousefi S., Simon H.U., Müller A.J, & Tacchini-Cottier F. (2015). Rapid Sequestration of Leishmania mexicana by Neutrophils Contributes to the Development of Chronic Lesion. PLoS Pathogens, 11(5), e1004929.
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8

Flow Cytometric Analysis of Cell Surface and Intracellular Markers

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For detection of surface markers and intracellular proteins, cells were harvested after trypsin treatment and resuspended in PBS supplemented with 2% FCS. Cells were pretreated with CD16/CD32 Fc-Block (eBioscience) for 20 min at 4°C. The following antibodies were used in concentrations according to instructions: SSEA-1-APC (51-8813-71), CD41-APC (17-0411-82), F4/80-APC (17-4801-80), CD11b-PE/eFluor 450 (12-0112-82/48-0112), CD14-FITC (11-0141-81), CD11c-PE-Cy7 (11-0141-81), Gr-1-eFluor450 (48-5931-82), CD45.1-PE-Cy7/APC (25-0453-81/17-0453), CD45.2-PE (12-0454-81), CD3-FITC (11-0031-81), B220-eFluor450 (48-0452-82), CD45-PE-Cy7/eFluor450 (25-0451-82/48-0451), CD200R-APC (17-5201), CD68-PE-Cy7 (25-0681) (all from eBioscience), SIGLEC-F-PE (552126, Becton Dickinson), Phospho-STAT5 (Tyr694) (C71E5) rabbit mAb (Alexa Fluor 647 conjugate) (9365, Cell Signaling), or Anti-Human/Mouse phospho-STAT5 (Y694) APC (17-9010, eBioscience). Fluorescence was measured using a BD FACSCalibur or BD LSR II and Fc-Block treated unstained sample was used as negative control.
Mucci A., Kunkiel J., Suzuki T., Brennig S., Glage S., Kühnel M.P., Ackermann M., Happle C., Kuhn A., Schambach A., Trapnell B.C., Hansen G., Moritz T, & Lachmann N. (2016). Murine iPSC-Derived Macrophages as a Tool for Disease Modeling of Hereditary Pulmonary Alveolar Proteinosis due to Csf2rb Deficiency. Stem Cell Reports, 7(2), 292-305.
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9

Hematopoietic Progenitor Cell Profiling

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Cell populations were identified and sorted on day 2 of blast culture based on surface markers. For experiments including hemogenic endothelium (HE) the floating and adherent cells were pooled. These cells were stained with KIT-APC (BD pharmingen), Tie2-PE (eBioscience) and CD41-PE-Cy7 (eBioscience) and analyzed on a Cyan ADP flow cytometer (Beckman Coulter) with data analysis using FlowJo, or sorted on a fluorescence associated cell sorting (FACS) Aria cell sorter (BD Biosciences). Progenitors matured in liquid culture were stained with CD11b-PE (eBioscience) and F4/80-APC (eBioscience).
Kellaway S.G., Keane P., Kennett E, & Bonifer C. (2020). RUNX1-EVI1 disrupts lineage determination and the cell cycle by interfering with RUNX1 and EVI1 driven gene regulatory networks. Haematologica, 106(6), 1569-1580.
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10

Multicolor Flow Cytometry of Myeloid Cells

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CD45 APC‐Cy7 (1:100, BioLegend, clone 30‐F11), MHC‐II PE (1:1,000, BioLegend, clone M5), CD11b PE‐Cy7 (1:100, BioLegend, clone M1/70), F4/80 APC (1:50, eBioscience, clone BM8), Ly6G PE (1:50, BioLegend), and 7AAD PerCP (2.5 μl/sample, Sigma).
Beck M.A., Fischer H., Grabner L.M., Groffics T., Winter M., Tangermann S., Meischel T., Zaussinger‐Haas B., Wagner P., Fischer C., Folie C., Arand J., Schöfer C., Ramsahoye B., Lagger S., Machat G., Eisenwort G., Schneider S., Podhornik A., Kothmayer M., Reichart U., Glösmann M., Tamir I., Mildner M., Sheibani‐Tezerji R., Kenner L., Petzelbauer P., Egger G., Sibilia M., Ablasser A, & Seiser C. (2021). DNA hypomethylation leads to cGAS‐induced autoinflammation in the epidermis. The EMBO Journal, 40(22), e108234.
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