Ccr2 pe cy7

Manufactured by BioLegend

Sourced in United States

CCR2-PE/Cy7 is a fluorochrome-conjugated antibody that binds to the C-C chemokine receptor type 2 (CCR2) expressed on the surface of cells. This product is designed for use in flow cytometry applications to identify and quantify CCR2-positive cells.

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CCR2-PE (2 citations) PE-Cy7-conjugated anti-mouse-CCR2 (SA203G11) (2 citations)

3 protocols using ccr2 pe cy7

Immune Cell Profiling in Blood Plasma

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Before collection of blood plasma, 25 µL blood was aliquoted from each sample and stained with antibodies against CD45-Alexa700 (103127), CD11b-FITC (101205), F4/80-PE (123109), Ly6C-Brilliant Violet 421 (128031), CCR2-PE/Cy7 (150611), CD3-APC (100235), CD4-PerCP/Cy5.5 (100539), CD8-Brilliant Violet 785 (100749), CD19-PE/Dazzle 594 (115553), and Ly6G-APC/Fire 750 (127651) (all purchased from BioLegend). Red blood cells (RBCs) were lysed with RBC lysis buffer (00-4333-57; eBioscience, Thermo Fisher Scientific), and samples were measured on the CytoFLEX Flow Cytometry System (Beckman Coulter). Results were analyzed using CytExpert acquisition and analysis software, version 2.4 (Beckman Coulter) and FlowJo analysis software (BD Biosciences).

van Dierendonck X.A., Vrieling F., Smeehuijzen L., Deng L., Boogaard J.P., Croes C.A., Temmerman L., Wetzels S., Biessen E., Kersten S, & Stienstra R. (2022). Triglyceride breakdown from lipid droplets regulates the inflammatory response in macrophages. Proceedings of the National Academy of Sciences of the United States of America, 119(12), e2114739119.

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Dexamethasone Modulates Macrophage Phenotype

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GM-CSF-differentiated macrophages were plated at 500K per well and treated with 0, 0.1, 1, or 10 uM dexamethasone in RPMI-1640 (Gibco) containing 10% FBS (Hyclone) for 24 hours, detached with Versene (Gibco), then analyzed immediately by flow cytometry. The following antibodies were used: CD40-BUV395, CD80-BV786, and HLA-DR/DP/DQ-FITC (BD Biosciences), HLA-ABC-Pacific blue, CD86-BV605, CD163-PerCPCy5.5, PD-L1-PE, and CCR2-PECy7 (BioLegend). For washout experiments, macrophages were treated with 0.1 uM dexamethasone for 24 hours, then standard RPMI-1640 with 10% FBS added for an additional 6 days prior to flow cytometry.

Moyes K.W., Davis A., Hoglund V., Haberthur K., Lieberman N.A., Kreuser S.A., Deutsch G.H., Franco S., Locke D., Carleton M.O., Gilbertson D.G., Simmons R., Winter C., Silber J., Gonzalez-Cuyar L.F., Ellenbogen R.G, & Crane C.A. (2018). Effects of tumor grade and dexamethasone on myeloid cells in patients with glioma. Oncoimmunology, 7(11), e1507668.

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Monocyte Phenotyping by Flow Cytometry

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All flow cytometry tests were performed on freshly prepared PBMCs and CD14 þ CD16 -monocytes. After blocking nonspecific antibody binding with human TruStain FcX (BioLegend, USA), the following monoclonal antibodies were used for flow cytometry: CD14-FITC, CD16-BV421, CCR1-PE, CCR2-PE/Cy7, CCR5-APC, CD11b-APC, CD18-PE (all from BioLegend, USA). We performed compensation using CompBeads (Becton Dickinson, USA). Flow cytometric analysis was performed on the CytoFLEX platform (Beckman Coulter, USA). The acquired data were analyzed by FlowJo software (Becton Dickinson, USA). Unstained monocytes were used as a negative control. Monocytes in PBMCs were identified as HLA-DR þ CD3 À CD19 -population with the unique forward and side-scatter properties.

Zhao X., Gu M., Xu X., Wen X., Yang G., Li L., Sheng P, & Meng F. (2020). CCL3/CCR1 mediates CD14⁺CD16^- circulating monocyte recruitment in knee osteoarthritis progression. Osteoarthritis and cartilage, 28(5).

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