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Three-color flow cytometric analysis of F4/80-Like Receptor expression on mouse bone marrow cells (left figures). Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD11b (Cat. No. 561690/553312) and FITC Hamster Anti-Mouse CD11c (Cat. No. 553801/557400/561045) antibodies and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602, Left Panels) or BD Horizon™ BV421 Rat Anti-Mouse F4/80-Like Receptor antibody (Cat. No. 563900; Right Panels). Two-color flow cytometric dot plots show the correlated expression patterns of CD11b (Upper Panels) or CD11c (Lower Panels) versus F4/80-Like Receptor (or Ig Isotype control staining) gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. Immunohistofluorescent analysis of F4/80-Like Receptor expression by cells within C57BL/6 mouse spleen (right figure). A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Rat Anti-Mouse F4/80-Like Receptor antibody (Cat. No. 563900, pseudo-colored green), BD Horizon™ BV480 Rat-anti-Mouse CD4 (Cat. No. 565634, pseudo-colored red), and Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557683, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
BD Horizon™ BV421 Rat Anti-Mouse F4/80-Like Receptor
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Recommended Assay Procedures
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).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
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- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
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Companion Products
The 6F12 antibody reacts with a 7-transmembrane-domain protein, which is similar to the F4/80 macrophage antigen of the EGF-TM7 protein family and is encoded by the Emr4 gene. The FIRE protein is expressed on myeloid cells with a denditic cell (DC) developmental potential, including subsets of DC and macrophages in the spleen and lymph nodes, most resident peritoneal macrophages, many peripheral blood monocytes, and a subpopulation of bone-marrow myeloid-cell progenitors. The protein is not detected on peripheral T and B lymphocytes, and it is down-regulated on thioglycollate-elicited peritoneal macrophages and on dendritic cells activated by GM-CSF, IFN-γ, anti-CD40, and LPS. Using soluble biotinylated fusion protein, a FIRE ligand was detected on a mouse IgG+ B lymphoma cell line (A20), but not on myeloid, fibroblast, or T-cell lines, suggesting that the FIRE protein may be involved in immunoregulatory interactions between antigen-presenting cells and B lymphocytes.
Development References (2)
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Caminschi I, Lucas KM, O'Keeffe MA, et al. Molecular cloning of F4/80-like-receptor, a seven-span membrane protein expressed differentially by dendritic cell and monocyte-macrophage subpopulations. J Immunol. 2001; 167(7):3570-3576. (Immunogen: Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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Stacey M, Chang GW, Sanos SL, et al. EMR4, a novel epidermal growth factor (EGF)-TM7 molecule up-regulated in activated mouse macrophages, binds to a putative cellular ligand on B lymphoma cell line A20. J Biol Chem. 2002; 277(32):29283. (Biology). View Reference
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