Invitrogen

F4/80 Monoclonal Antibody (BM8), PE-eFluor 610, eBioscience™

Product Image Gallery

F4/80 Antibody — Figure 2 Functionality of bone marrow (BM)-derived macrophages after spraying. ( a ) Flow cytometric analysis of cell viability by propidium iodide (PI) staining of unsprayed and sprayed BM-derived macrophages (Left: representative images, right: quantification of n = 3, mean +- SEM). ( b ) Papenheim staining of cytospin preparations of sprayed and unsprayed BM-derived macrophages (scale bar: 20 um) ( c ) Flow cytometric analysis of CD45.1, CD11b and F4/80 expression of unsprayed and sprayed BM-derived macrophages. Overlays of representative histograms; grey filled line: isotype, red/blue line: surface marker of unsprayed or sprayed macrophages. Lower panels: Delta mean fluorescent intensity (MFI) of surface marker expression of unsprayed and sprayed BM-derived macrophages (delta MFI was calculated by subtracting isotype (negative) MFI from surface marker (positive) MFI; n = 3, mean +- SEM). ( d ) Phagocytic activity of unsprayed and sprayed BM-derived macrophages. Upper panels: representative fluorescent microscopy images of BM-derived macrophages 2 hours after incubation with pHRodo E. coli BioParticles (scale bar: 200 um). Lower panel: Representative flow cytometric analysis (grey filled line: cells without E. coli BioParticles, red line: unsprayed cells with E. coli BioParticles, blue line: sprayed cells with E. coli BioParticles) and quantification of n = 3 (mean +- SEM). ( e ) Analysis of MHC-II surface marker expression by flow cytometry of unsprayed and spraye

F4/80 Antibody in Flow Cytometry (Flow) — Staining of mouse resident peritoneal exudate cells with Anti-Mouse CD11b APC (Product # 17-0112-82) and 0.5 µg of Rat IgG2a K Isotype Control PE-eFluor® 610 (Product # 61-4321-82) (left) or 0.5 µg of Anti-Mouse F4/80 Antigen PE-eFluor® 610 (right). Cells in the large scatter population were used for analysis.

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

1 µg/test

Published Applications

Flow Cytometry (Flow)

See 65 publications below

Miscellaneous PubMed (MISC)

See 1 publication below

Immunohistochemistry (Frozen) (IHC (F))

See 1 publication below

Product Specifications

Species Reactivity

Mouse

Published species

Fish, Hamster, Human, Mouse

Host / Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), PE-eFluor 610, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

BM8

Conjugate

PE-eFluor® 610

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_2574612

Product Specific Information

Description: The BM8 monoclonal antibody reacts with mouse F4/80 antigen, an approximately 160 kDa surface receptor. It belongs to the EGF-TM7 family of proteins. As such it contains seven EGF-like domains on its extracellular N-terminus, seven transmembrane spanning sequences, and an intracellular C-terminal domain showing homology to other TM7 superfamily members. The F4/80 antigen is expressed by a majority of mature macrophages and is one of the best markers for this population of cells. However, other cell types, such as peritoneal eosinophils, Langerhans cells, and some other dendritic cell subtypes, have been reported to express this antigen as well. Expression of F4/80 commences during early myeloid development in vivo and can be upregulated on BM cells stimulated in vitro with M-CSF. Some populations of macrophages, especially in the lymphoid microenvironment, may be devoid of F4/80.

Applications Reported: This BM8 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This BM8 antibody has been tested by flow cytometric analysis of mouse resident peritoneal exudate cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

F4/80 antigen is a 160 kDa cell surface glycoprotein that is a member of the EGF TM7 family of proteins and shares 68% overall amino acid identity with human EGF module containing mucin like hormone receptor 1 (EMR1). Expression of F4/80 is heterogeneous and is reported to vary during macrophage maturation and activation. The F4/80 antigen is expressed on a wide range of mature tissue macrophages including Kupffer cells, Langerhans, microglia, macrophages located in the gut lamina propria, peritoneal cavity, lung, thymus, bone marrow stroma and macrophages in the red pulp of the spleen. F4/80 expression has also been reported on a subpopulation of dendritic cells but is absent from macrophages located in T cell areas of the spleen and lymph node. The ligands and biological functions of the F4/80 antigen have not yet been determined but recent studies suggest a role for F4/80 in the generation of efferent CD8+ve regulatory T cells.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Adhesion G protein-coupled receptor E1; Cell surface glycoprotein F4/80; DD7A5-7; EGF-like module containing, mucin-like, hormone receptor-like sequence 1; EGF-like module receptor 1; EGF-like module-containing mucin-like hormone receptor-like 1; EGF-TM7; EMR1 hormone receptor; F4/80; F480; Gpf480; Ly71; lymphocyte antigen 71; TM7LN3

Gene Aliases: Adgre1; DD7A5-7; EGF-TM7; Emr1; F4/80; Gpf480; Ly71; TM7LN3

UniProt ID: (Mouse) Q61549

Entrez Gene ID: (Mouse) 13733

Molecular Function: G-protein coupled receptor antibacterial response protein defense/immunity protein receptor