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Analysis of CD206 Expression by Thioglycolate-elicited Peritoneal Exudate Cells (Thio PECs) Surface CD206 Expression - Panel 1: Thio PECs were stained with Alexa Fluor™ 647 Rat Anti-Mouse CD107b antibody (Cat. No. 564843) and with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; Top Plot) or BD Horizon™ BUV395 Rat Anti-Mouse CD206 antibody (Cat. No. 568817/568818; Bottom Plot) at 0.5 μg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD206 (or Ig Isotype control staining) versus CD107b was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Surface and Intracellular CD206 Expression - Panel 2: Thio PECs were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained as previously described at 0.5 µg/test. The bivariate plots showing the correlated expression of CD206 (or Ig Isotype control staining) versus CD107b were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific and were generated in separate experiments.
BD Horizon™ BUV395 Rat Anti-Mouse CD206
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Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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Companion Products
The Y17-505 monoclonal antibody specifically binds to CD206 which is also known as the Macrophage mannose receptor (MMR, MR) or Mannose receptor, C type 1 (Mrc1). CD206 is a type I transmembrane glycoprotein of approximately 175 kDa that belongs to the C-type lectin superfamily. It is expressed at the cell surface and intracellularly by macrophages, Langerhans cells, dendritic cells, and endothelial cells within hepatic and lymphoid tissues. This pattern recognition receptor binds to endogenous and microbial glycoconjugates containing mannose, fucose, or N-acetylglucosamine through its C-type lectin-like carbohydrate recognition domains (CRD). CD206 also contains a cysteine-rich domain that enables binding to sulfated carbohydrate antigens. This receptor enables macrophages and other specialized cells to maintain tissue homeostasis as well as to internalize microbes or their components by phagocytosis or endocytosis. CD206 thus plays important roles in mediating innate immunity, eg, enabling phagocytosis, as well as in processing and presenting antigens for the generation and expression of adaptive immunity. Moreover, CD206 has been associated with leucocyte homing and cancer cell metastasis.
Development References (6)
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Akbarshahi H, Menzel M, Posaric Bauden M, Rosendahl A, Andersson R. Enrichment of murine CD68+ CCR2+ and CD68+ CD206+ lung macrophages in acute pancreatitis-associated acute lung injury. PLoS ONE. 2012; 7(10):e42654. (Biology). View Reference
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Burgdorf S, Lukacs-Kornek V, Kurts C. The mannose receptor mediates uptake of soluble but not of cell-associated antigen for cross-presentation. J Immunol. 2006; 176(11):6770-6776. (Biology). View Reference
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Marttila-Ichihara F, Turja R, Miiluniemi M, et al. Macrophage mannose receptor on lymphatics controls cell trafficking. Blood. 2008; 112(1):64-72. (Biology). View Reference
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McKenzie EJ, Taylor PR, Stillion RJ, et al. J Immunol. 2007; 178(8):4975-4983. (Biology). View Reference
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Rybalko V, Hsieh PL, Merscham-Banda M, Suggs LJ, Farrar RP. The Development of Macrophage-Mediated Cell Therapy to Improve Skeletal Muscle Function after Injury.. PLoS One. 2015; 10(12):e0145550. (Biology). View Reference
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Zamze S, Martinez-Pomares L, Jones H, et al. Recognition of bacterial capsular polysaccharides and lipopolysaccharides by the macrophage mannose receptor. J Biol Chem. 2002; 277(44):41613-41623. (Biology). View Reference
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