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PerCP-Cy5.5 Rat Anti-Mouse F4/80
PerCP-Cy5.5 Rat Anti-Mouse F4/80
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 for splenocytes or DRAQ7™-negative for PEC) monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ 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 PE Rat Anti-CD11b antibody (Cat. No. 553311/557397/561689) and with either PerCP-Cy5.5 Rat IgG2a Isotype Control (Cat. No. 550765; Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse F4/80 antibody (Cat. No. 567202; Right Plot) at 0.5 µg/test. DAPI 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 BD Horizon™ BV421 Rat Anti-Mouse CD117 antibody (Cat. No. 566290/562609) and either PerCP-Cy5.5 Rat IgG2a Isotype Control (Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse F4/80 antibody (Right Plot) at 0.5 µg/test. A solution containing DRAQ7™ (Cat. No. 564904) was added to cells right before analysis.
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 for splenocytes or DRAQ7™-negative for PEC) monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ 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 PE Rat Anti-CD11b antibody (Cat. No. 553311/557397/561689) and with either PerCP-Cy5.5 Rat IgG2a Isotype Control (Cat. No. 550765; Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse F4/80 antibody (Cat. No. 567202; Right Plot) at 0.5 µg/test. DAPI 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 BD Horizon™ BV421 Rat Anti-Mouse CD117 antibody (Cat. No. 566290/562609) and either PerCP-Cy5.5 Rat IgG2a Isotype Control (Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse F4/80 antibody (Right Plot) at 0.5 µg/test. A solution containing DRAQ7™ (Cat. No. 564904) was added to cells right before analysis.
Product Details
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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
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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. 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.
  4. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  5. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  9. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  10. An isotype control should be used at the same concentration as the antibody of interest.
567202 Rev. 1
Antibody Details
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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.

567202 Rev. 1
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
567202 Rev.1
Citations & References
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Development References (7)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
View All (7) View Less
567202 Rev. 1

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