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BV421 Rat Anti-Mouse F4/80
BV421 Rat Anti-Mouse F4/80

Two-color flow cytometric analysis of mouse F4/80 expression.

       Panel 1 - Spleen. 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 Alexa Fluor® 647 Rat Anti-Mouse CD11b antibody (Cat. No. 557686) and either BD Horizon™ BV421 Rat IgG2a Isotype Control (Cat. No. 562602; Left Plot) or BD Horizon BV421 Rat Anti-Mouse F4/80 antibody (Cat. No. 565411; Right Plot). The two-color flow cytometric contour 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.

       Panel 2 - Peritoneal. C57BL/6 mouse peritoneal cells were preincubated with Mouse BD Fc Block™ and stained with APC Rat Anti-Mouse CD117 (Cat. No. 553356/561074) and BV421 Rat Anti-Mouse F4/80 antibodies. The two-color contour plot showing the correlated expression of F4/80 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.

Image analysis of F4/80 expression in mouse spleen.

       Panel 3 - Spleen. A section from a paraformaldehyde-fixed and frozen normal C57BL/6 mouse spleen was stained with BV421 Rat Anti-Mouse F4/80 (pseudo-colored green), Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 (Cat. No. 557683; pseudo-colored red), and BD Horizon™ BV480 Anti-CD3 (pseudo-colored aqua) antibodies. The image was captured on a standard Epifluorescence microscope using 392/23nm excitation and 430/24nm emission.

Two-color flow cytometric analysis of mouse F4/80 expression.

       Panel 1 - Spleen. 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 Alexa Fluor® 647 Rat Anti-Mouse CD11b antibody (Cat. No. 557686) and either BD Horizon™ BV421 Rat IgG2a Isotype Control (Cat. No. 562602; Left Plot) or BD Horizon BV421 Rat Anti-Mouse F4/80 antibody (Cat. No. 565411; Right Plot). The two-color flow cytometric contour 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.

       Panel 2 - Peritoneal. C57BL/6 mouse peritoneal cells were preincubated with Mouse BD Fc Block™ and stained with APC Rat Anti-Mouse CD117 (Cat. No. 553356/561074) and BV421 Rat Anti-Mouse F4/80 antibodies. The two-color contour plot showing the correlated expression of F4/80 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.

Image analysis of F4/80 expression in mouse spleen.

       Panel 3 - Spleen. A section from a paraformaldehyde-fixed and frozen normal C57BL/6 mouse spleen was stained with BV421 Rat Anti-Mouse F4/80 (pseudo-colored green), Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 (Cat. No. 557683; pseudo-colored red), and BD Horizon™ BV480 Anti-CD3 (pseudo-colored aqua) antibodies. The image was captured on a standard Epifluorescence microscope using 392/23nm excitation and 430/24nm emission.

Product Details
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BD Horizon™
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), Bioimaging (Tested During Development)
0.2 mg/ml
13733
AB_2734779
Aqueous buffered solution containing BSA and ≤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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. An isotype control should be used at the same concentration as the antibody of interest.
565411 Rev. 2
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.

 

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565411 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565411 Rev.2
Citations & References
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Development References (5)

  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. 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
  3. 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
  4. 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
  5. 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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565411 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.