M0M-CSF (2×106/mL) were treated with CS (0.2 µM) or vehicle (0.1% DMSO) for 48 hours. Then, cells were stained in PBS pH 7.4 containing 0.5% bovine serum albumin, 2 mM EDTA, and 0.1% sodium azide by Zombie Aqua™ Fixable Viability Kit (Biolegend, San Diego, CA) for 5 minutes at 4°C to determine cell viability. Non-specific antibody binding was blocked by using mouse serum (10 minutes at 4°C) prior to staining by the following fluorochrome-labeled antibodies (20 minutes, 4°C): FITC anti-human CD14 (clone M5E2, #555397, BD Biosciences, San Jose, CA), APC-H7 anti-human CD80 (clone L307.4, #561134, BD Biosciences), PE-Cy7 anti-human CD54 (clone HA58, #353115, Biolegend), PE anti-human CD163 (clone GHI/61, #556018, BD Biosciences), and APC anti-human CD206 (clone 19.2, #550889, BD Biosciences) to determine M1 and M2 surface marker expression using a LSRFortessaTM cell analyzer (BD Biosciences), and data were analyzed using FlowJo X Software (BD Biosciences).
Fitc anti human cd14 clone m5e2
Manufactured by BD
Sourced in United States
FITC anti-human CD14 (clone M5E2) is a fluorescently labeled monoclonal antibody that binds to the CD14 cell surface marker on human cells. CD14 is a glycosylphosphatidylinositol-anchored protein that plays a role in the innate immune response. This antibody can be used for the identification and enumeration of CD14-positive cells by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Apc h7 anti human cd80 clone l307, by BD (3 mentions)
Pe cy7 anti human cd54 clone ha58, by BioLegend (3 mentions)
Pe anti human cd163 clone ghi 61, by BD (3 mentions)
Apc anti human cd206 clone 19, by BD (3 mentions)
Zombie aqua fixable viability kit, by BioLegend (3 mentions)
Lsrfortessa cell analyzer, by BD (3 mentions)
3 protocols using fitc anti human cd14 clone m5e2
1
Macrophage Polarization Analysis
2
Macrophage Immunophenotyping and Viability
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Macrophages were stained in PBS pH 7.4 containing 0.5% BSA, 2 mmol/L EDTA and 0.1% sodium azide by Zombie Aqua™ Fixable Viability Kit (Biolegend, San Diego, CA, USA) to determine cell viability (5 min, 4 °C) and by fluorochrome-labelled antibodies (20 min, 4 °C): FITC anti-human CD14 (clone M5E2, #555397, BD Bioscience, San Jose, CA), APC-H7 anti-human CD80 (clone L307.4, #561134, BD Bioscience), PE-Cy7 anti-human CD54 (clone HA58, #353115, Biolegend), PE anti-human CD163 (clone GHI/61, #556018, BD Bioscience), APC anti-human CD206 (clone 19.2, #550889, BD Bioscience) to determine M1 and M2 surface marker expression using a LSRFortessa™ cell analyzer (BD Bioscience), and data were analyzed using FlowJo X Software (BD Bioscience)21 (link).
3
M1/M2 Macrophage Polarization Assay
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M0M-CSF MDM were treated with TWG (1 µg/mL) or vehicle (0.1% DMSO) for 48 h. Then, cells were stained in PBS pH 7.4 containing 0.5% BSA, 2 mM EDTA and 0.1% sodium azide by Zombie Aqua™ Fixable Viability Kit (Biolegend, San Diego, CA, USA) for 5 min at 4 °C to determine cell viability. Non-specific antibody binding was blocked by using mouse serum (10 min at 4 °C) prior to staining by the following fluorochrome-labelled antibodies (20 min, 4 °C): FITC anti-human CD14 (clone M5E2, #555397, BD Bioscience, San Jose, CA, USA), APC-H7 anti-human CD80 (clone L307.4, #561134, BD Bioscience), PE-Cy7 anti-human CD54 (clone HA58, #353115, Biolegend, Koblenz, Germany), PE anti-human CD163 (clone GHI/61, #556018, BD Biosciences, Heidelberg, Germany), APC anti-human CD206 (clone 19.2, #550889, BD Bioscience) to determine M1 and M2 surface marker expression using LSRFortessaTM cell analyzer (BD Bioscience), and data were analyzed using FlowJo X Software (BD Bioscience).
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