Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Bacteria - Escherichia coli, Nematode - Caenorhabditis elegans

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Radioimmunoassay

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2B Clone # Cl:A3-1

Format

BSA Free
View all F4/80 Primary Antibodies »

Product Specifications

Immunogen

Thioglycollate stimulated peritoneal macrophages from C57BL/6 mice

Localization

Cell surface

Marker

Macrophage Marker

Clonality

Monoclonal

Host

Rat

Isotype

IgG2B

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.

Scientific Data Images for F4/80 Antibody (Cl:A3-1) - BSA Free

Immunohistochemistry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

F4-80-Antibody-CI-A3-1-Immunohistochemistry-NB600-404-img0052.jpg
Immunocytochemistry/ Immunofluorescence: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunocytochemistry/ Immunofluorescence: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunocytochemistry/Immunofluorescence: F4/80 Antibody (CI-A3-1) [NB600-404] - Lymph node stained for macrophages, red and B cells CD79b, green; nuclei are stained with DAPI, blue.
Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB600-404] - A surface stain was performed on RAW 246.7 cells with F4/80 antibody (CI-A3-1) NB600-404AF488 (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 10 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Immunohistochemistry-Paraffin: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry-Paraffin: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

F4-80-Antibody-CI-A3-1-Immunohistochemistry-Paraffin-NB600-404-img0050.jpg
Immunocytochemistry/ Immunofluorescence: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunocytochemistry/ Immunofluorescence: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunocytochemistry/Immunofluorescence: F4/80 Antibody (CI-A3-1) [NB600-404] - Analysis of monocytes cultured in M-CSF using F4/80 antibody. Primary antibody dilution: 1:200. Overnight incubation in RT. Image from verified customer review.
Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404] - Surface staining of F4/80 in CT26 colorectal carcinoma tumor model. Using Allophycocyanin conjugated version of the antibody (NB600-404APC). Image from verified customer review
Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) [NB600-404] - Frozen mouse spleen with F4/80 anti-mouse antibody.
Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) [NB600-404] - Mouse spleen stained with A: Rat anti Mouse. B: CI-A3-1 clone preincubated with 2 molar excess of human anti Idiotypic.
Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB600-404] - Staining of J774 cells with F4/80 antibody.
Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) - BSA Free [NB600-404]

Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB600-404] - Staining of mouse peritoneal macrophages with clone CI-A3-1 F4/80 antibody.
Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB600-404]
F4/80 Antibody (Cl:A3-1)

Immunohistochemistry: F4/80 Antibody (Cl:A3-1) [NB600-404] -

Immunohistochemistry: F4/80 Antibody (Cl:A3-1) [NB600-404] - Lymphocyte interaction inside the DC harboring scaffoldsDC scaffolds (beDC) or scaffolds only, placed in tumor bearing mice or DC-scaffold placed in normal mice were recovered two weeks post-implantation & processed for immunohistochemistry to demonstrate various cell types. (A) Relative size of the biomatrices harvested from various mice groups as depicted in the figure (far left panel). n = 3–5. The dotted line in ‘gross specimen’ differentiates the biomatrix from the reactive host-tissue. Representative images of biomatrix sections stained by the respective antibodies as indicated in the figure. The bar in the ‘gross structure’ is equivalent to 1 mm, whereas for other panels is equal to 100 μm. (B) Snapshots from Supplementary Movie S2 showing lymphocyte (green) movement & interaction with DCs (red) inside the biomatrix. The start point of the observation was set as the zero time point. (C) Lymphocyte division (white arrows) inside the biomatrix in the vicinity of DC (red). Representative snapshot from Supplementary Movie S3. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27223090), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
F4/80 Antibody (Cl:A3-1) - BSA Free

Western Blot: F4/80 Antibody (Cl:A3-1) - BSA Free [NB600-404] -

MFD alters immune signaling in adipose tissue during (a) acute and (b) chronic Mtb infection. Immunoblot analysis of immune cells (CD4, CD8, and F4/80) and inflammatory markers (IFN gamma, TNF alpha, IL-6, and IL-10) in the WAT lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during acute infection (30 DPI) and chronic (3 months) post-infection is shown. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36676177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
F4/80 Antibody (Cl:A3-1) - BSA Free

Western Blot: F4/80 Antibody (Cl:A3-1) - BSA Free [NB600-404] -

MFD alters the level of infiltrated immune cells and inflammatory markers in the lungs during chronic Mtb infection. Immunoblot analysis of (a) immune cells (CD4+, CD8+, and F4/80+) and (b) inflammatory markers (IFN gamma, TNF alpha, and IL-6) in the lung lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during chronic (90 DPI) infection. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36676177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
F4/80 Antibody (Cl:A3-1) - BSA Free

Western Blot: F4/80 Antibody (Cl:A3-1) - BSA Free [NB600-404] -

MFD alters immune signaling in adipose tissue during (a) acute and (b) chronic Mtb infection. Immunoblot analysis of immune cells (CD4, CD8, and F4/80) and inflammatory markers (IFN gamma, TNF alpha, IL-6, and IL-10) in the WAT lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during acute infection (30 DPI) and chronic (3 months) post-infection is shown. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36676177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for F4/80 Antibody (Cl:A3-1) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:50-1:100

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500RadioImmunoassay 1:100-1:2000

Radioimmunoassay

1:100-1:2000

Western Blot

1:100-1:2000
Application Notes
This product requires pretreatment of paraffin sections prior to staining. Proteinase K is recommended for tissues fixed for less than 24 hours. Citrate buffer pH 6.0 is recommended for tissues fixed for more than 24 hours. E. coli reactivity reported in scientific literature (PMID:31181736).

Reviewed Applications

Read 2 reviews rated 4 using NB600-404 in the following applications:

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 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 short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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

Entrez Gene IDs

13733 (Mouse)

Gene Symbol

ADGRE1

UniProt

Q61549

Additional F4/80 Products

Product Documents for F4/80 Antibody (Cl:A3-1) - BSA Free

Certificate of Analysis

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

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Product Specific Notices for F4/80 Antibody (Cl:A3-1) - BSA Free

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.

Citations for F4/80 Antibody (Cl:A3-1) - BSA Free

Customer Reviews for F4/80 Antibody (Cl:A3-1) - BSA Free (2)

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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • F4/80 Antibody (CI-A3-1)
    Name: Mozhdeh Sojoodi
    Application: Immunocytochemistry
    Sample Tested: Bone Marrrow Derived Macrophages
    Species: Mouse
    Verified Customer | Posted 02/07/2022
    Monocytes cultured in M-CSF are positive for F4/80
    1:200 Dilution Overnight incubation in RT
    F4/80 Antibody (Cl:A3-1) - BSA Free NB600-404
  • Name: Naiche Adler
    Application: Immunohistochemistry-Paraffin
    Sample Tested: mouse lymph node FFPE
    Species: Mouse
    Verified Customer | Posted 03/26/2015
    mouse lymph node
    F4/80 Antibody (Cl:A3-1) - BSA Free NB600-404

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Showing  1 - 2 of 2 reviews Showing All

Protocols

View specific protocols for F4/80 Antibody (Cl:A3-1) - BSA Free (NB600-404):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for F4/80 Antibody (Cl:A3-1) - BSA Free

Showing  1 - 4 of 4 FAQs Showing All

  • Q: Are the F4/80/EMR1 antibodies validated in Simple Western?

    A: No, F4/80/EMR1 antibodies are not tested in Simple Western at this moment.

  • Q: Does F4/80/EMR1 antibodies come in lyophilized form?

    A: Yes, we carry 2 F4/80 antibodies in lyophilized form: MAB5580 and NBP1-06975

  • Q: I'm hoping to use F4/80 as a macrophage marker in skeletal muscle using IHC-Fr but I haven't seen this done successfully (and heard people have tried without much luck). Do you know of any validation that has been done on that tissue?

    A: We have not validated NB600-404 on skeletal muscle specifically, and we haven't yet had any customer feedback regarding this antibody being used on skeletal muscle. If you are interested in using this antibody for your testing, and have any trouble getting it to work for you, please let us know. We do offer a guarantee on our products for the species and applications listed, and if we can't get this product working for you, we will compensate you for the purchase.

  • Q: What is the theoretical molecular weight for F4/80/EMR1 antibodies?

    A: The TMW of F4/80/EMR1 antibodies is approximately 160 kDa.

  • Q: Are the F4/80/EMR1 antibodies validated in Simple Western?

    A: No, F4/80/EMR1 antibodies are not tested in Simple Western at this moment.

  • Q: Does F4/80/EMR1 antibodies come in lyophilized form?

    A: Yes, we carry 2 F4/80 antibodies in lyophilized form: MAB5580 and NBP1-06975

  • Q: I'm hoping to use F4/80 as a macrophage marker in skeletal muscle using IHC-Fr but I haven't seen this done successfully (and heard people have tried without much luck). Do you know of any validation that has been done on that tissue?

    A: We have not validated NB600-404 on skeletal muscle specifically, and we haven't yet had any customer feedback regarding this antibody being used on skeletal muscle. If you are interested in using this antibody for your testing, and have any trouble getting it to work for you, please let us know. We do offer a guarantee on our products for the species and applications listed, and if we can't get this product working for you, we will compensate you for the purchase.

  • Q: What is the theoretical molecular weight for F4/80/EMR1 antibodies?

    A: The TMW of F4/80/EMR1 antibodies is approximately 160 kDa.

  • Q: Are the F4/80/EMR1 antibodies validated in Simple Western?

    A: No, F4/80/EMR1 antibodies are not tested in Simple Western at this moment.

  • Q: Does F4/80/EMR1 antibodies come in lyophilized form?

    A: Yes, we carry 2 F4/80 antibodies in lyophilized form: MAB5580 and NBP1-06975

  • Q: I'm hoping to use F4/80 as a macrophage marker in skeletal muscle using IHC-Fr but I haven't seen this done successfully (and heard people have tried without much luck). Do you know of any validation that has been done on that tissue?

    A: We have not validated NB600-404 on skeletal muscle specifically, and we haven't yet had any customer feedback regarding this antibody being used on skeletal muscle. If you are interested in using this antibody for your testing, and have any trouble getting it to work for you, please let us know. We do offer a guarantee on our products for the species and applications listed, and if we can't get this product working for you, we will compensate you for the purchase.

  • Q: What is the theoretical molecular weight for F4/80/EMR1 antibodies?

    A: The TMW of F4/80/EMR1 antibodies is approximately 160 kDa.

  • Q: Are the F4/80/EMR1 antibodies validated in Simple Western?

    A: No, F4/80/EMR1 antibodies are not tested in Simple Western at this moment.

  • Q: Does F4/80/EMR1 antibodies come in lyophilized form?

    A: Yes, we carry 2 F4/80 antibodies in lyophilized form: MAB5580 and NBP1-06975

  • Q: I'm hoping to use F4/80 as a macrophage marker in skeletal muscle using IHC-Fr but I haven't seen this done successfully (and heard people have tried without much luck). Do you know of any validation that has been done on that tissue?

    A: We have not validated NB600-404 on skeletal muscle specifically, and we haven't yet had any customer feedback regarding this antibody being used on skeletal muscle. If you are interested in using this antibody for your testing, and have any trouble getting it to work for you, please let us know. We do offer a guarantee on our products for the species and applications listed, and if we can't get this product working for you, we will compensate you for the purchase.

  • Q: What is the theoretical molecular weight for F4/80/EMR1 antibodies?

    A: The TMW of F4/80/EMR1 antibodies is approximately 160 kDa.

Showing  1 - 4 of 4 FAQs Showing All
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