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 BD Horizon™ BUV395 Rat Anti-Mouse CD11b antibody (Cat. No. 563553/565976) and with either Biotin Rat IgG2a Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.06 µg/test. The cells were washed and counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown in this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of F4/80 expression on mouse peritoneal cells. C57BL/6 mouse peritoneal exudate cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with BD Horizon™ BV421 Rat Anti-Mouse CD117 antibody (Cat. No. 562609/566290) and with either Biotin Rat IgG2a Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.06 µg/test. The cells were washed and counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown in this Technical Data Sheet are not lot specific.
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 BD Horizon™ BUV395 Rat Anti-Mouse CD11b antibody (Cat. No. 563553/565976) and with either Biotin Rat IgG2a Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.06 µg/test. The cells were washed and counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown in this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of F4/80 expression on mouse peritoneal cells. C57BL/6 mouse peritoneal exudate cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with BD Horizon™ BV421 Rat Anti-Mouse CD117 antibody (Cat. No. 562609/566290) and with either Biotin Rat IgG2a Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse F4/80 antibody (Cat. No. 566787; Right Plot) at 0.06 µg/test. The cells were washed and counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown in 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)
0.5 mg/ml
13733
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
PE Streptavidin RUO
Size
0.5 mg
Cat No.
554061
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BUV395 Rat Anti-CD11b RUO
Size
50 µg
Cat No.
563553
BUV395 Rat Anti-CD11b RUO
Size
25 µg
Cat No.
565976
BV421 Rat Anti-Mouse CD117 RUO
Size
50 µg
Cat No.
562609
567893 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.
567893 Rev. 1
Format Details
Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
567893 Rev.1
Citations & References
Development References (5)
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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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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
567893 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.
