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Two-color flow cytometric analysis of F4/80 expression on mouse splenocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with APC Rat Anti-Mouse CD11b antibody (Cat. No. 553312/561690) and either BD Horizon™ BV711 Rat IgG2a Isotype Control (Cat. No. 563047; Left Plot) or BD Horizon BV711 Rat Anti-Mouse F4/80 antibody (Cat. No. 565612; Right Plot). The two-color flow cytometric dot plot showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometric analyses were performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of F4/80 expression on mouse peritoneal cells. C57BL/6 mouse peritoneal cells were preincubated with Mouse BD Fc Block™ and stained with APC Rat Anti-Mouse CD117antibody (Cat. No. 553356/561074) and either BD Horizon™ BV711 Rat IgG2a Isotype Control (Left Plot) or BD Horizon BV711 Rat Anti-Mouse F4/80 antibody (Right Plot). The two-color bot plot showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD117 was derived from gated events with the light-scatter characteristics of viable peritoneal cells. Flow cytometric analyses were performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BV711 Rat Anti-Mouse F4/80
BD Horizon™ BV711 Rat Anti-Mouse F4/80
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Cy is a trademark of Amersham Biosciences Limited.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The T45-2342 monoclonal antibody recognizes the mouse F4/80 antigen which is also known as EGF-like module-containing mucin-like hormone receptor-like 1 (EMR1). F4/80 is a 160 kDa glycoprotein that belongs to the EGF-TM7 family of seven-transmembrane spanning cell surface molecules. It is expressed on the surface of granulocytes and a wide range of mature tissue macrophages including, Kupffer cells, splenic red pulp macrophages, microglia, gut lamina propria macrophages, and Langerhans cells. F4/80 expression has also been reported on subpopulations of dendritic cells. F4/80 expression is heterogeneous and may be increased during inflammatory responses as observed in various mouse models of colitis, diabetes and brain injury.
The antibody was conjugated to BD Horizon BV711 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (6)
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Austyn JM., and Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981; 10:805-815. (Biology). View Reference
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Bodhankar S, Lapato A, Chen Y, Vandenbark AA, Saugstad JA, Offner H. Role for microglia in sex differences after ischemic stroke: importance of M2.. Metab Brain Dis. 2015. (Clone-specific: Flow cytometry). View Reference
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Gordon S, Hamann J, Lin HH, Stacey M. F4/80 and the related adhesion-GPCRs. Eur J Immunol. 2011; 41(9):2472-2476. (Biology). View Reference
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Krüger T, Benke D, Eitner F, et al. Identification and functional characterization of dendritic cells in the healthy murine kidney and in experimental glomerulonephritis. J Am Soc Nephrol. 2004; 15(3):613-621. (Biology). View Reference
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Leenen PJ, Radosević K, Voerman JS, et al. Heterogeneity of mouse spleen dendritic cells: in vivo phagocytic activity, expression of macrophage markers, and subpopulation turnover.. J Immunol. 1998; 160(5):2166-73. (Biology). View Reference
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McKnight AJ, Macfarlane AJ, Dri P, Turley L, Willis AC, Gordon S. Molecular cloning of F4/80, a murine macrophage-restricted cell surface glycoprotein with Homology to the G-protein-linked transmembrane & hormone receptor family. J Biol Chem. 1996; 271:486. (Biology). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.