F4 80 pe cy7 clone bm8

Manufactured by BioLegend

Sourced in United States

F4/80-PE-CY7 (clone BM8) is a fluorescently-labeled antibody reagent used for the detection and analysis of F4/80 positive cells in flow cytometry applications.

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Lab products found in correlation

Cell stimulation cocktail and protein transport inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Cd16 cd32, by BD (1 mentions) Ly6g bv421, by BioLegend (1 mentions) Il 4 bv605, by BioLegend (1 mentions) Zombie nir fixable viability kit, by BioLegend (1 mentions) Cd11c pe, by BioLegend (1 mentions) Ifn γ pe cy7, by BioLegend (1 mentions) Il 17a bv510, by BioLegend (1 mentions) Il 10 pe jes5 16e3, by BD (1 mentions) Foxp3 transcription buffer set, by Thermo Fisher Scientific (1 mentions) Attune nxt software, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Cd4 fitc, by BD (1 mentions) Attune nxt acoustic focusing cytometer, by Thermo Fisher Scientific (1 mentions) Thiazole orange, by Merck Group (1 mentions) Ifn γ apc, by BioLegend (1 mentions) Tnf α fitc, by BD (1 mentions) Ter119 pe clone ter 119, by BD (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions)

3 protocols using f4 80 pe cy7 clone bm8

Multiparameter Flow Cytometry of Immune Cells

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Cells in BALF were stained with CD45-FITC (clone 30-F11), CD11b-APC (clone M1/70), CD11c-PE (clone N418), F4/80-PE-CY7 (clone BM8), and Ly6G-BV421 (clone 1A8, BioLegend) in the presence of an Fc blocker (CD16/CD32, BD Biosciences). A total of 2 × 10⁶ single lung cells were stimulated with cell stimulation cocktail and protein transport inhibitor cocktail (Cat#00-4970-03 and #00-4980-93, eBioscience) in IMDM medium supplemented with 10% FCS, 2 mM L-glutamine, 100 U/ml penicillin-streptomycin for 4 h in a 37°C incubator with 5% CO₂. The cells were incubated with a Zombie NIR™ Fixable Viability Kit (Cat#423105, BioLegend) at room temperature for 15 min and then stained with antibodies to extracellular antigens in the presence of Fc blocker at 4°C for 25 min. Intracellular and nuclear staining was performed with the Foxp3/Transcription buffer set (Cat# 00-5523-00, eBiosciences). Data were acquired using LSRFortessa and FACSDiva software (BD Biosciences) or Attune NxT 3 L-BRV and Attune NxT software (Thermo Fisher Scientific) and analyzed using FlowJo 10.7.2 (TreeStar, Ashland). The following antibodies were used: CD4-FITC (clone GK1.5), TCR-BV421 (clone H57-597), Foxp3-AF647 (clone MF23), IL-10-PE (JES5-16E3, BD Biosciences), IFN-γ-PE-Cy7 (clone XMG1.2), IL-4-BV605 (clone 11B11), IL-17a-BV510 (clone TC11-18H10.1, Biolegend), and IL-13-PE-eFluor 610 (eBiosciences).

Li J.D, & Yin J. (2023). Interleukin-10-alveolar macrophage cell membrane-coated nanoparticles alleviate airway inflammation and regulate Th17/regulatory T cell balance in a mouse model. Frontiers in Immunology, 14, 1186393.

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Alveolar Macrophage Polarization and Particle Uptake

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Isolated alveolar macrophages were polarized and plated in 6-well plates for 24-hr at 37°C (as described above). After 24-hr the phenotype media was aspirated, cells washed with PBS and provided RPMI media (described above) supplemented with either vehicle (control volume of 100% EtOH) or DHA (25 μM) for 24-hr at 37°C. Particles (MWCNT or SiO₂, 50 μg/ml) and LPS (20 ng/ml) were added to the cells for 4-hr at 37°C. Particle uptake was determined by flow cytometry using the Attune® NxT Acoustic Focusing Cytometer (Life Technologies; Carlsbad, CA, USA) the Flow Cytometry Core at the University of Montana by utilizing changes in side scatter (SSC) and anti-mouse antibody F4/80-PE/Cy7 (clone BM8, Biolegend; San Diego, CA, USA) to identify alveolar macrophages. Data analysis was performed in FlowJo 10.6.

Fletcher P., Hamilton RF J.r., Rhoderick J.F., Pestka J.J, & Holian A. (2020). Docosahexaenoic acid impacts macrophage phenotype subsets and phagolysosomal membrane permeability with particle exposure. Journal of toxicology and environmental health. Part A, 84(4), 152-172.

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Flow Cytometry Panel for Immune Cell Analysis

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Flow cytometry was performed using a BD Accuri C6 flow cytometer (BD Biosciences), and data were analyzed in FlowJo v10. Cells were sorted on a Beckman Coulter MoFlo Astrios. Flow antibodies: c-Kit Alexa Fluor 647 (clone 2B8, 105818, BioLegend), Sca-1 phycoerythrin (PE)-Cy7 (clone D7, BDB558162, BD Biosciences), Ter119 PE (clone Ter119, BDB553673, BD Biosciences), CD71 FITC (fluorescein isothiocyanate) (clone C2, BDB553266, BD Biosciences), F4/80 PE-Cy7 (clone BM8, 123112, BioLegend), CD172a allophycocyanin (APC) (clone P84, BDB560106, BD Biosciences), FITC TNF-α (506304, BD Biosciences), FITC IL-1β (IC4013F, R&D Systems), and APC IFN-γ (505809, BioLegend). Working concentration for all flow cytometry antibodies was 1:200. For reticulocytes, whole blood was stained with thiazole orange (390062, Sigma-Aldrich) at room temperature for 1 hour and then analyzed by flow cytometry.

Bennett L.F., Liao C., Quickel M.D., Yeoh B.S., Vijay-Kumar M., Hankey-Giblin P., Prabhu K.S, & Paulson R.F. (2019). Inflammation induces stress erythropoiesis through heme-dependent activation of SPI-C. Science signaling, 12(598), eaap7336.

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