BD Horizon™ RB705 Rat Anti-Mouse F4/80
Clone T45-2342 (RUO)
Multicolor flow cytometric analysis of F4/80 expression on viable Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The leukocytes were then stained with APC Rat Anti-CD11b antibody (Cat. No. 553312) and with either BD Horizon™ RB705 Rat IgG2a, κ Isotype Control (Cat. No. 570262; Left Plot) or BD Horizon™ RB705 Rat Anti-Mouse F4/80 antibody (Cat. No. 570288/570289; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-negative) monocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of F4/80 expression on viable Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The leukocytes were then stained with APC Rat Anti-CD11b antibody (Cat. No. 553312) and with either BD Horizon™ RB705 Rat IgG2a, κ Isotype Control (Cat. No. 570262; Left Plot) or BD Horizon™ RB705 Rat Anti-Mouse F4/80 antibody (Cat. No. 570288/570289; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-negative) monocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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)
0.2 mg/ml
13733
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- 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 observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB705 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570262
APC Rat Anti-CD11b RUO
Size
0.1 mg
Cat No.
553312
DAPI Solution RUO
Size
1 mg
Cat No.
564907
570288 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.
570288 Rev. 1
Format Details
RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
RB705
Blue 488 nm
498 nm
707 nm
570288 Rev.1
Citations & References
View product citations for antibody "570288" on CiteAb
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
570288 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.