BD Horizon™ RB780 Rat Anti-Mouse F4/80
Clone T45-2342 (RUO)
Multicolor flow cytometric analyses of F4/80 expression. The bivariate pseudocolor density plots showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD11b (Mouse Splenocytes; Top Plots) or CD117 (Mouse Peritoneal Exudate Cells; Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) monocytes. 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.
Top Plots - 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 BD Horizon™ BUV395 Rat Anti- CD11b antibody (Cat. No. 563553) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse F4/80 antibody (Cat. No. 569223/569224; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis.
Bottom Plots - Mouse peritoneal exudate cells (PEC). C57BL/6 Mouse PEC were similarly pretreated with Mouse BD Fc Block™, stained with PE Rat Anti-Mouse CD117 antibody (Cat. No. 553355/561075) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse F4/80 antibody (Right Plot) at 0.5 µg/test followed by DAPI Solution and analyzed.
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Multicolor flow cytometric analyses of F4/80 expression. The bivariate pseudocolor density plots showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD11b (Mouse Splenocytes; Top Plots) or CD117 (Mouse Peritoneal Exudate Cells; Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) monocytes. 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.
Top Plots - 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 BD Horizon™ BUV395 Rat Anti- CD11b antibody (Cat. No. 563553) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse F4/80 antibody (Cat. No. 569223/569224; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis.
Bottom Plots - Mouse peritoneal exudate cells (PEC). C57BL/6 Mouse PEC were similarly pretreated with Mouse BD Fc Block™, stained with PE Rat Anti-Mouse CD117 antibody (Cat. No. 553355/561075) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse F4/80 antibody (Right Plot) at 0.5 µg/test followed by DAPI Solution and analyzed.
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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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.
Development References (7)
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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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Ito F, Ku AW, Bucsek MJ, et al. Immune Adjuvant Activity of Pre-Resectional Radiofrequency Ablation Protects against Local and Systemic Recurrence in Aggressive Murine Colorectal Cancer.. PLoS ONE. 2015; 10(11):e0143370. (Clone-specific: Flow cytometry). 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
Please refer to Support Documents for Quality Certificates
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.
Please refer to Support Documents for Quality Certificates
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.