Anti f4 80 bv421

Manufactured by BioLegend

Anti-F4/80-BV421 is a fluorochrome-conjugated antibody that binds to the F4/80 antigen, which is expressed on the surface of mature mouse macrophages. This product is intended for use in flow cytometry applications.

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Lab products found in correlation

Anti cd11b pe, by BioLegend (3 mentions) Anti cd16 32, by BioLegend (2 mentions) Collagenase d, by Merck Group (2 mentions) Recombinant human il 4, by Thermo Fisher Scientific (1 mentions) Recombinant mouse il 6, by Thermo Fisher Scientific (1 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions) U0126, by Merck Group (1 mentions) Arginase 1, by Cell Signaling Technology (1 mentions) Stat3, by ABclonal (1 mentions) Mouse m csf, by Thermo Fisher Scientific (1 mentions) P erk1 2, by ABclonal (1 mentions) Polyinosinic polycytidylic acid poly, by InvivoGen (1 mentions) Mouse ifn β, by PBL Assay Science (1 mentions) Stat6, by ABclonal (1 mentions) Stat1, by ABclonal (1 mentions) Live dead fixable violet dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Anti cd4 apc, by Thermo Fisher Scientific (1 mentions) Anti cd4 pe cy7, by BioLegend (1 mentions) Anti cd45 bv711, by BioLegend (1 mentions) Zombie nir, by BioLegend (1 mentions)

Spelling variants of this product

F4/80 (349 citations) Anti-F4/80 (96 citations) F4/80-BV421 (20 citations) BV421 labeled anti-F4/80 (3 citations)

9 protocols using anti f4 80 bv421

Polarization of Murine and Human Macrophages

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Polyinosinic–polycytidylic acid (poly(I:C)) was purchased from InvivoGen. Mouse IFNβ (#12405-1) was from PBL. Mouse M-CSF (#315-02), mouse recombinant IL-6 (#315-05), mouse recombinant IL-4 (#214-14), human recombinant M-CSF (#300-25-2), human recombinant IL-6 (#200-06), human recombinant IL-4 (#200-04) were from PeproTech. The ELISA kits for mIL-6 (#88-7064-22) and mIL-4 (#88-7044-22) were from eBioscience.
Mouse IL-4-neutralizing antibodies were from Bio-x cell (#BE0045), mouse IL-6-neutralizing antibody (#504512) was from Biolegend. The ERK1/2 inhibitor U0126 (#U120) was from Sigma. SHP099 (#HY-1003881) was from MCE. The NE-PER Nuclear and Cytoplasmic Extraction kit (#78833) was from Thermo Fisher. Primary antibodies for Western blot were purchased from Cell Signaling Technology (arginase-1, #93668; pSTAT6-Y641, #565543; STAT6, #93623; pSTAT3-Tyr705, #9145; STAT3, #9139; STAT1, #14994; Erk1/2, #4695; pErk1/2, #4370), and Genscript (Actin, #A00730), Abclonal (GFP, #AE012). Antibodies for Flow Cytometry were purchased form Biolegend: anti-CD16/32 (#101330), AF488-anti-Ly6C (#128022), BV421-anti-F4/80 (#123132), AF647-anti-CD206 (#141712).

Yang L., Guo P., Wang P., Wang W, & Liu J. (2023). IL-6/ERK signaling pathway participates in type I IFN-programmed, unconventional M2-like macrophage polarization. Scientific Reports, 13, 1827.

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Comprehensive Tumor Immune Cell Profiling

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Tumors were chopped into small pieces that were then transferred into gentleMACS Tubes (MACS Miltenyi Biotec), containing 10 mL of DMEM media and 1 mg/mL collagenase D (Sigma-Aldrich, COLLD-RO Roche, #11088866001). The tubes were placed on a gentleMACS Dissociator (MACS Miltenyi Biotec, #130-095-937) using the program 37_m_TDK2. After incubation, cells were filtered using 70 µm cell strainer and recovered by centrifugation. Cells were stained for live/dead with either LIVE/DEAD Fixable Violet Dead Cell Stain Kit, for 405 nm excitation (Thermo Fisher, cat#L34963) or Zombie NIR (BioLegend, cat#423105) then stained with a cocktail of surface mAbs Panel 1: BV711 anti-CD45 (BioLegend, cat#103147), PE anti-NK1.1 (BioLegend, cat#108707) and PE/Cy7 anti-CD8 (eBioscience, cat# 25-0083), APC anti-CD4 (eBioscience, cat#14-0042-81), BV421 anti-F4/80 (BioLegend, cat#123137) and PE/Dazzle 594 anti-CD183 (CXCR3) (BioLegend, cat#155914); Panel 2: BV711 anti-CD45 (BioLegend, cat#103147), APC anti-IFNg (BioLegend, cat#505810) and PE/Dazzle 594 anti-T-bet (BioLegend, cat#644828), PE/Cy7 anti-CD4 (BioLegend, cat#100422). After 30 min of staining, cells were washed and samples were run on FACS Symphony Cytometer (BD Biosciences). FlowJo V.10 was used for the analysis, cells were manually gated on size and granularity. Dead cells and doublets were excluded, and CD45 + cells were selected.

Fitzgerald A.A., Wang S., Agarwal V., Marcisak E.F., Zuo A., Jablonski S.A., Loth M., Fertig E.J., MacDougall J., Zhukovsky E., Trivedi S., Bhatia D., O'Neill V, & Weiner L.M. (2021). DPP inhibition alters the CXCR3 axis and enhances NK and CD8+ T cell infiltration to improve anti-PD1 efficacy in murine models of pancreatic ductal adenocarcinoma. Journal for Immunotherapy of Cancer, 9(11), e002837.

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Flow Cytometric Phenotyping of Macrophages

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Monocyte-derived macrophages or BMDMs were harvested and washed in cold PBS. Cells were incubated in Fc-block reagent (BD biosciences) and fixable viability dye eFluor 780 (Invitrogen) for 15 min at 4°C in cold PBS. Monocyte-derived macrophages were washed once and stained with the following antibodies: anti-CD62L PerCp-Cy5.5 (BD biosciences), anti-CCR2 APC (R and D Systems), anti-F4/80 efluor450 (eBioscience), anti-CSF1R BV711 (Biolegend), anti-Ly6G BUV395 (BD biosciences), anti-CD11b BUV737 (BD biosciences), anti-MHCII(I-A/I-E) PE (eBioscience) and anti-Ly6C PE-Cy7 (eBioscience). BMDMs were washed once and stained with the following antibodies: anti-CD45 FITC (eBioscience), anti-F4/80 BV421 (Biolegend), anti-CD11b BUV395 (BD biosciences). Stained cells were analyzed on by flow cytometry using a BD FACS CANTO instrument. Loss of eGFP or CD45 was assessed by gating on live F4/80 + macrophages using FlowJo X.

Lim J., Park H., Heisler J., Maculins T., Roose-Girma M., Xu M., Mckenzie B., van Lookeren Campagne M., Newton K, & Murthy A. (2019). Autophagy regulates inflammatory programmed cell death via turnover of RHIM-domain proteins. eLife, 8, e44452.

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Profiling Lung-Infiltrating Immune Cells

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When the lung-infiltrating cells were analyzed, anti-CD45-APC-Cy7 (BioLegend) were intravenously injected (3 μg/mouse) before euthanasia to exclude the blood-circulating immune cells. Cells from lung or spinal cord tissues were prepared by enzymatic digestion with 2.5 mg/ml collagenase D (Sigma-Aldrich) and 0.1 mg/ml DNase I (Sigma-Aldrich) for 30 min at 37°C. For neutrophils, monocytes, macrophages, and CD4⁺ T cell staining, the cells or tissues were stained with anti-Ly6c-AF488 (BioLegend), anti-CD45.2-PE (BioLegend), anti-CD11c-PE (BioLegend), anti-CD11b-PerCP (BioLegend), anti-Gr-1-APC (eBioscience), anti-CD4-APC (TONBO), and anti-F4/80-BV421 (BioLegend).

Miyashita Y., Kouwaki T., Tsukamoto H., Okamoto M., Nakamura K, & Oshiumi H. (2021). TICAM-1/TRIF associates with Act1 and suppresses IL-17 receptor–mediated inflammatory responses. Life Science Alliance, 5(2), e202101181.

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Analyzing Nanoparticle Uptake in Lung Cells

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The ALI mice were treated with Cy5.5-labeled NPs for 22 h. The cell suspensions in BALF were stained with anti-CD11b-AF488, anti-CD11c-APC, anti-F4/80-BV421, and anti-Ly6G-BV785 antibodies (BioLegend), and analyzed using CytoFLEX flow cytometer (Beckman Coulter, Inc.). Data were analyzed with FlowJo software X.

Zhai Z., Ouyang W., Yao Y., Zhang Y., Zhang H., Xu F, & Gao C. (2022). Dexamethasone-loaded ROS-responsive poly(thioketal) nanoparticles suppress inflammation and oxidative stress of acute lung injury. Bioactive Materials, 14, 430-442.

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Peritoneal Macrophage Isolation and Culture

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Peritoneal lavage was performed with 5 ml of freshly-prepared room temperature 0.2 μm sterile-filtered PBS containing 2 mM EDTA and 1 % FBS, per animal. All cells for each sex and age group were collected, pooled, washed, and resuspended in cold DMEM supplemented with 10 % FBS and 2 mM EDTA. Blocking was performed by adding 20 μg/ml anti-CD16/32 (Biolegend #101301) and incubated for 15 minutes on ice in dark. Subsequently, 1.24 μg/ml anti-F4/80 BV421 (Biolegend #123137) and anti-CD11b PE (Biolegend #101208) were added followed by an incubation in dark for 30 min. Cells were washed and resuspended in 500 μl cold DMEM supplemented with 10 % FBS and 2 mM EDTA on ice. ToPro3 (ThermoFisher Scientific #T3605) was added at a dilution of 1:1000 prior to sorting to gate out dead cells. CD11b^hi F4/80^hi cells were sorted into 1 ml DMEM supplemented with 20 % FBS. ~0.5E6 cells were subsequently cultured in DMEM supplemented with 10 % FBS in 6-well tissue culture dishes overnight before proceeding to LPS stimulation.

Babagana M., Oh K.S., Chakraborty S., Pacholewska A., Aqdas M, & Sung M.H. (2021). Hedgehog dysregulation contributes to tissue-specific inflammaging of resident macrophages. Aging (Albany NY), 13(15), 19207-19229.

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Apoptosis Analysis of Boron Carbide Cytotoxicity

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The RAW264.7 and J774A.1 cells (0.5 × 10⁵ cells/well for 24 h incubation and 0.25 × 10⁵ cells/well for 72 h in 24-well plates), JAWS II cells (1 × 10⁵ cells/well for 24 h incubation and 0.5 × 10⁵ cells/well for 72 h in 24-well plates) were incubated with boron carbide preparations at concentrations 10, 50, 100 and 200 µg/ml, and BMDM (1.2 × 10⁵ cells/well for both times) at concentrations 10, 50 and 100 µg/ml for 24 and 72 h. After this time, cells were harvested, suspended in a binding buffer and centrifuged (10 min, 100 × g, 4 °C). Next, cells were stained with Annexin V protein conjugated with APC fluorochrome (Becton Dickinson) for 15 min at room temperature. BMDM were additionally stained with anti-F4/80 BV421 (BioLegend) for 45 min at 4 °C. To assess the percentage of dead cells, 10 µg/ml propidium iodide (PI; Invitrogen) was added. The cells were analyzed using flow cytometer LSRFortessa with Diva Software (Becton Dickinson). Scheme of flow cytometry analysis was performed using NovoExpress software 1.3.0 (ACEA Biosciences, Inc.). Apoptosis assays were performed in two independent experiments, each in triplicate.

Wróblewska A., Szermer-Olearnik B., Szczygieł A., Węgierek-Ciura K., Mierzejewska J., Kozień D., Żeliszewska P., Kruszakin R., Migdał P., Pędzich Z, & Pajtasz-Piasecka E. (2024). Macrophages as carriers of boron carbide nanoparticles dedicated to boron neutron capture therapy. Journal of Nanobiotechnology, 22, 183.

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Murine Immune Cell Profiling

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To block the nonspecific binding of antibodies to Fcγ receptors, isolated single cell suspensions were incubated first with anti-CD16/32 antibody (101302, Biolegend) at 4°C for 10 min. Subsequently, the cells were incubated with a mixture of antibodies at 4°C for 25 min. Anti-CD11b-PE (Cat: 101208, Biolegend), anti-Gr1-FITC (Cat: 108406, Biolegend), anti-F4/80-BV421 (Cat: 123132, Biolegend) and anti-CD206-Alexa674 (Cat: 565250, BD Pharmingen) were used for flow cytometric analysis. The obtained results were expressed as the percent. Flow cytometric data was analyzed using official FlowJo software.

Wu Q., Xu R., Zhang K., Sun R., Yang M., Li K., Liu H., Xue Y., Xu H, & Guo Y. (2023). Characterization of early myocardial inflammation in ischemia-reperfusion injury. Frontiers in Immunology, 13, 1081719.

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Flow Cytometric Analysis of Immune Cell Subsets

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The following fluorescent-labeled monoclonal antibodies and staining reagents are used according to the manufacturer's specifications: anti-APC-CY7-live (BioLegend), anti-BV510-CD45 (BioLegend), anti-F4/80-BV421 (BioLegend), anti-CD11B-PE CY7 (BioLegend), anti-CD206-APC (BioLegend), anti-CD80-percp/Cy5.5 (BioLegend), anti-LY6C-APC (BioLegend), anti-CD11B-PE (BioLegend), anti-LY6G-PE CY7 (BioLegend), anti-CD3-PE CY7 (BioLegend), anti-CD4-BV421 (BioLegend), anti-CD8-percp/CY5.5 (BioLegend). The cells were proceeded by FACS Calibur flow cytometry and analyzed by the FlowJo software.

Li W., Yu J., Jin B., Zhang H, & Zhang J. (2021). Protective Effects of Aminooxyacetic Acid on Colitis Induced in Mice with Dextran Sulfate Sodium. BioMed Research International, 2021, 1477345.

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