Anti cd206 fitc clone dcn228

Manufactured by Miltenyi Biotec

Anti-CD206 FITC clone DCN228 is a fluorescent-labeled monoclonal antibody that binds to the CD206 antigen, also known as the mannose receptor. CD206 is a cell surface receptor expressed on specific cell types, including macrophages and dendritic cells. This product can be used for the identification and characterization of CD206-positive cells in various applications, such as flow cytometry and immunohistochemistry.

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Lab products found in correlation

Cell dissociation buffer, by Thermo Fisher Scientific (1 mentions) Live dead aqua, by Thermo Fisher Scientific (1 mentions) Gallios flow cytometer, by Beckman Coulter (1 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) Human fcr blocking reagent, by Miltenyi Biotec (1 mentions) Human ab serum, by Lonza (1 mentions) Em grade, by Electron Microscopy Sciences (1 mentions) Lsrfortessa, by BD (1 mentions)

Close spelling variants of this product

CD206-FITC DCN228 CD206

2 protocols using anti cd206 fitc clone dcn228

Phenotypic Analysis of M1 and M2 Macrophages

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Macrophages were harvested using cell dissociation buffer (Invitrogen) for flow cytometric analyses. Cells were stained in LiveDead Aqua (Invitrogen) and then incubated with human FcR blocking reagent (Miltenyi). For purity analyses post CD14⁺ isolation, cells were stained with anti-CD14 FITC clone TÜK4 (Miltenyi) and anti-CD3 PE clone REA613 (Miltenyi). For M1 and M2 macrophage phenotypic analyses, cells were stained with anti-CD206 FITC clone DCN228 (Miltenyi), anti-CD163 PEDazzle clone GHI/61 (BioLegend), anti-CD80 BV421 clone 2D10 (BioLegend), and anti-CD86 AF700 clone FUN-1 (BD Pharmingen). Isotype controls were also stained in parallel. Data for Figs 1 and 2 were collected on a Gallios flow cytometer (Beckman Coulter) and subsequent data on a Cytoflex flow cytometer (Beckman Coulter). FlowJo software version 10.4.2 (Tree Star) was used to analyze the results. Cells were gated to exclude dead cells before differential expression analyses.

Gerrick K.Y., Gerrick E.R., Gupta A., Wheelan S.J., Yegnasubramanian S, & Jaffee E.M. (2018). Transcriptional profiling identifies novel regulators of macrophage polarization. PLoS ONE, 13(12), e0208602.

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Monocyte Phospho-STAT Profiling by Flow Cytometry

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Flow cytometry experiments were performed with purified monocytes. For surface staining, cells were stained (20 minutes at 4°C) in PBS + 10% human AB serum (Lonza) + 0.05% NaN₃ (Staining buffer, SB). For phosphoprotein staining, cells were rested in cIMDM-5 for 1 hour and stimulated with the indicated cytokines for 15 minutes. Then, cells were fixed with paraformaldehyde (EM-grade, Electron Microscopy Sciences) (final concentration 1.5%) and permeabilized with absolute ice-cold methanol as previously described [60 ]. Cells were stained (30 minutes at room temperature) in SB. The following antibodies were used: anti-CD123 FITC (clone AC145, dilution 1:20), anti-CD206 FITC (clone DCN228, dilution 1:20) (Miltenyi Biotec), anti-phospho-STAT5 AlexaFluor 647 (clone 47, dilution 1:20) and anti-phospho-STAT6 AlexaFluor 647 (clone 18, dilution 1:20) (BD Biosciences). Samples were acquired on a BD LSRFortessa and analyzed using FACSDiva Software. Data are expressed as percentage of positive cells and median fluorescence intensity (MFI) of positive cells. Gating strategies for surface staining and phosphoprotein staining are shown in Supporting Information Fig. 13 and Fig. 14, respectively.

Borriello F., Longo M., Spinelli R., Pecoraro A., Granata F., Staiano R.I., Loffredo S., Spadaro G., Beguinot F., Schroeder J, & Marone G. (2015). IL-3 synergizes with basophil-derived IL-4 and IL-13 to promote the alternative activation of human monocytes. European journal of immunology, 45(7), 2042-2051.

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