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 PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either APC Rat IgG2a Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. 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 (7-AAD negative) monocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of F4/80 expression on mouse peritoneal cells. 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 PE Rat Anti-Mouse CD117 (Cat. No. 553355/561075) and either APC Rat IgG2a Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The two-color dot plot showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD117 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD negative) peritoneal cells. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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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 PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either APC Rat IgG2a Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. 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 (7-AAD negative) monocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of F4/80 expression on mouse peritoneal cells. 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 PE Rat Anti-Mouse CD117 (Cat. No. 553355/561075) and either APC Rat IgG2a Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The two-color dot plot showing the correlated expression of F4/80 (or Ig Isotype control staining) versus CD117 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD negative) peritoneal cells. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Gpf480; F480; Emr1; Ly71; DD7A5-7; EGF-TM7; TM7LN3
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Mouse F4/80 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
13733
AB_2869866
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.
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.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- 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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
APC Rat IgG2a κ Isotype Control RUO
Size
0.1 mg
Cat No.
553932
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566787 Rev. 1
Antibody Details
T45-2342
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.
566787 Rev. 1
Format Details
APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC
Red 627-640 nm
651 nm
660 nm
566787 Rev.1
Citations & References
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
566787 Rev. 1
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.
