F4/80 Antibody (CI:A3-1)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-61613

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP2-61613
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Key Product Details

Species Reactivity

Mouse

Applications

Validated:

Flow Cytometry, Immunohistochemistry

Cited:

IF/IHC

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rat IgG1 kappa Clone # CI:A3-1

Concentration

1 mg/ml

View all F4/80 Antibodies »

Product Specifications

Immunogen

This recombinant F4/80 Antibody (CI:A3-1) was developed against Thioglycollate stimulated peritoneal macrophages of mouse origin.

Clonality

Monoclonal

Host

Rat

Isotype

IgG1 kappa

Theoretical MW

160 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

Recombinant monoclonal antibody to F4/80. Manufactured using Recombinant Platform with variable regions (i.e. specificity) from the hybridoma CI:A3-1 (recombinant version).

Scientific Data Images for F4/80 Antibody (CI:A3-1)

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613] - Mouse (Mus musculus) splenocytes were labelled ex vivo with a commercially available APC-labelled anti-F4/80 antibody and APE labelled anti-CD11b antibody and subject to flow-cytometry analysis (A), in which a small subpopulation of F4/80-CD11b positive cells may be observed. Subsets of commerical anti-F4/80 antibody-labelled splenocytes were also subsequently incubated with unlabelled versions of either the rat (Rattus norvegicus) IgG2b chimeric version (B) or mouse IgG2A chimeric (C) version of CI:A3-1. Loss of the F4/80-CD11b positive subpopulation may be observed, demonstrating displacement of the commercial antibody and the specificity of CI:A3-1
Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613] - Figure 1 murine bone marrow-derived macrophages (BMDMs) were pre-blocked with rat anti-mouse CD16 & CD32 (clone FCR-4G8) and stained with non-recombinant anti-F4/80 conjugated to Alexa Fluor 647 (AF647), all commercially available from competitors.
Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613]

Flow Cytometry: F4/80 Antibody (CI:A3-1) [NBP2-61613] - Figure 2 BMDMs were stained with recombinant anti-F4/80 or isotype control (anti-fluorescein [4-4-20 (enhanced)] IgG2b. Whilst in Figure 2 the highest fluorescence signal is seen with the recombinant anti-F4/80 IgG2 (the isotype of the original hybridoma-derived antibody), the isotype control IgG2b shows considerable signal overlap, indicative of binding of the antibody to Fc-receptors. This illustrates the importance of isotype controls in such experiments when using conventional antibody formats particularly when Fc-blocking reagents are incompatible with the system used due to reactivity with the secondary antibody.
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Applications for F4/80 Antibody (CI:A3-1)

Application
Recommended Usage

Flow Cytometry

1:10-1:1000

Immunohistochemistry

1:10-1:500

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

Background: F4/80

F4/80 antigen is a cell surface glycoprotein that is the murine homolog of human EGF-like module-containing mucin-like hormone receptor-like 1 (EMR1) (1,2). Specifically, F4/80 (EMR1) is one of five members belonging to the adhesion G-coupled receptor (ADGRE) subfamily, also known as the epidermal growth factor seven transmembrane (EGF-TM7) receptors (1,2). F4/80 is expressed by the myeloid lineage including in eosinophils, monocytes, macrophages, and myeloid dendritic cells (1,2). The molecular structure of F4/80 contains an EGF-like extracellular domain and a seven transmembrane motif, with a theoretical molecular weight of 160 kDa (1).

Depending on factors such as differentiation, activation, and location, the level of F4/80 expression can vary between cell-types (1,2). For example, peritoneal macrophages are subdivided based on morphology and expression with large peritoneal macrophages having high levels of F4/80 and small peritoneal macrophages having low F4/80 expression (1). Furthermore, splenic macrophages are classified based on location (red pulp, marginal zone, and white pulp) and F4/80 expression level varies within these populations as well; the red pulp macrophages have high levels of F4/80 and CD68 by comparison (1). Originally, the F4/80 monoclonal antibody was generated for a macrophage-specific surface antigen, however F4/80 staining is also observed in other hematopoietic cell populations, thus other immune cell markers should be used in conjunction with F4/80 to distinguish between different populations (1). Though the full extent of F4/80 ligands and biological functions remains elusive, studies have suggested that F4/80 is critical for macrophage-natural killer cell interaction and cytokine release (1,2). Additionally, a study of anterior chamber-associated immune deviation (ACAID) indicated a possible role for F4/80 in the generation of regulatory T cells (1,2).

References

1. Dos Anjos Cassado A. (2017). F4/80 as a Major Macrophage Marker: The Case of the Peritoneum and Spleen. Results and Problems in Cell Differentiation. https://doi.org/10.1007/978-3-319-54090-0_7

2. Hamann, J., Hsiao, C. C., Lee, C. S., Ravichandran, K. S., & Lin, H. H. (2016). Adhesion GPCRs as Modulators of Immune Cell Function. Handbook of Experimental Pharmacology. https://doi.org/10.1007/978-3-319-41523-9_15

Alternate Names

DD7A5-7, Gpf480, Ly71

Gene Symbol

Adgre1

Additional F4/80 Products

Product Documents for F4/80 Antibody (CI:A3-1)

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for F4/80 Antibody (CI:A3-1)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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