Overview

  • Product name
    Anti-F4/80 antibody [CI:A3-1]
    See all F4/80 primary antibodies
  • Description
    Rat monoclonal [CI:A3-1] to F4/80
  • Specificity
    This antibody recognises the mouse F4/80 antigen, a 160kD glycoprotein expressed by murine macrophages. Abcam does not recommend this product for IHC-P.
  • Tested applications
    Suitable for: WB, IP, RIA, IHC-Fr, Flow Cyt, ICC, IHC-FoFr, ICC/IF, IHC-Rmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Thioglycollate stimulated peritoneal macrophages from C57/BL mice

  • Positive control
    • Mouse spleen and macrophages ICC/IF: Raw264.7 cells.
  • General notes

    Please note we cannot guarantee IHC-P, however should you use this application for this product then please use a specific protocol which can be found in the protocols section of our datasheet here.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.60
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: 0.164% Sodium phosphate, 1.45% Sodium chloride, 1% BSA
  • Concentration information loading...
  • Purity
    IgG fraction
  • Purification notes
    Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
  • Clonality
    Monoclonal
  • Clone number
    CI:A3-1
  • Myeloma
    NS1
  • Isotype
    IgG2b
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6640 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Use under non reducing condition. Detects a band of approximately 160 kDa (predicted molecular weight: 102 kDa).

(predicted molecular weight of precursor protein: 102 kDa; protein is heavily glycosylated). Block in 5% milk for 1 hour.

IP Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
IHC-Fr Use a concentration of 10 µg/ml.
Flow Cyt 1/10 - 1/50.

ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC Use at an assay dependent concentration.
IHC-FoFr Use a concentration of 10 µg/ml.
ICC/IF Use a concentration of 5 - 10 µg/ml.
IHC-R Use a concentration of 10 µg/ml.

Target

  • Function
    Orphan receptor involved in cell adhesion and probably in cell-cell interactions specifically involving cells of the immune system. May play a role in regulatory T-cells (Treg) development.
  • Tissue specificity
    Expression is restricted to eosinophils.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 2 family. Adhesion G-protein coupled receptor (ADGR) subfamily.
    Contains 6 EGF-like domains.
    Contains 1 GPS domain.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ADGRE1 antibody
    • Adhesion G protein coupled receptor E1 antibody
    • Adhesion G protein-coupled receptor E1 antibody
    • AGRE1_HUMAN antibody
    • Cell surface glycoprotein EMR1 antibody
    • Cell surface glycoprotein F4/80 antibody
    • DD7A5 7 antibody
    • Egf like module containing mucin like hormone receptor like 1 antibody
    • Egf like module containing mucin like hormone receptor like sequence 1 antibody
    • EGF like module receptor 1 antibody
    • EGF TM7 antibody
    • EGF-like module receptor 1 antibody
    • EGF-like module-containing mucin-like hormone receptor-like 1 antibody
    • EGFTM7 antibody
    • EMR 1 antibody
    • EMR1 hormone receptor antibody
    • Gpf480 antibody
    • Ly71 antibody
    • Lymphocyte antigen 71 antibody
    • TM7LN3 antibody
    see all

Anti-F4/80 antibody [CI:A3-1] images

  • Paraffin-embedded mouse spleen tissue stained for F4/80 using ab6640 at 1/8000 dilution in immunohistochemical analysis.

    See Abreview

  • Paraffin-embedded mouse liver tissue stained for F4/80 using ab6640 at 1/50 dilution in immunohistochemical analysis.

    See Abreview

  • ab6640 stained Raw 264.7 cells. The cells were 100% methanol fixed for 5 minutes at room temperatureand then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6640 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150165 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-F4/80 antibody [CI:A3-1] (ab6640) at 1/1000 dilution

    Lane 1 : Mouse macrophage lysates
    Lane 2 : Mouse hepatocyte lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP labelled donkey anti-rat at 1/5000 dilution

    Predicted band size : 102 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab6640 staining F4/80 in mouse spleen tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde and blocked with 5% serum for 45 minutes at 21°C. The sample was incubated with primary antibody (1/100 in PBS + 5% FBS) at 4°C for 16 hours. An Alexa Fluor® 488-conjugated goat anti-rat IgG (H+L) polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab6640 staining F4/80 in mouse bone marrow cells by immunocytochemistry/ immunofluorescence. Cells were formaldehyde fixed and permeabilized in 0.2% Triton X-100 prior to blocking in 2% BSA for 30 minutes at 20°C. The primary antibody was diluted 1/200 and incubated with the sample for 9 hour at 4°C. Alexa fluor® 488 goat polyclonal to rat Ig, diluted 1/200, was used as the secondary antibody.

    See Abreview

  • Flow cytometry analysis of J774 cells labelling F4/80 (red) with ab6640 at a dilution of 1/10 followed by streptavidin FITC secondary at 1/100 dilution. The blue line shows J774 cells stained with Rat anti mouse isotype control. 

  • Flow cytometry analysis of peritoneal macrophages labelling F4/80 (red) with ab6640 at a dilution of 1/10. The blue line shows J774 cells stained with Rat anti mouse isotype control. 

  • Flow cytometry analysis of J774 cells labelling F4/80 (red) with ab6640 at a dilution of 1/50 followed by goat anti rat IgG FITC secondary antibody at 1/100 dilution. The blue line shows J774 cells stained with Rat anti mouse isotype control. 

  • ab6640 staining F4/80 (red) in Mouse intestine tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/10µg/ml in PBS + 10% serum) for 16 hours at 4°C. An Alexa Fluor® 555-conjugated Donkey anti-rat IgG polyclonal (1/1000) was used as the secondary antibody. Blue - nuclei.

    See Abreview

References for Anti-F4/80 antibody [CI:A3-1] (ab6640)

This product has been referenced in:
  • Xue N  et al. Chlorogenic acid inhibits glioblastoma growth through repolarizating macrophage from M2 to M1 phenotype. Sci Rep 7:39011 (2017). IHC-Fr ; Mouse . Read more (PubMed: 28045028) »
  • Hughes KR  et al. Bifidobacterium breve reduces apoptotic epithelial cell shedding in an exopolysaccharide and MyD88-dependent manner. Open Biol 7:N/A (2017). IHC-P ; Mouse . Read more (PubMed: 28123052) »

See all 268 Publications for this product

Product Wall

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Bone marrow-derived macrophages)
Permeabilization
Yes - Triton X-100
Specification
Bone marrow-derived macrophages
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 14 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.3% triton x100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

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Verified customer

Submitted Mar 24 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Heart)
Permeabilization
Yes - Tween-20
Specification
Embryonic Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 11 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted Oct 04 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrite buffer, pH6.0
Permeabilization
No
Specification
spleen
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 19 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Jaculus jaculus Tissue sections (spleen)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: proteaseK
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Mai Tran

Verified customer

Submitted Jul 12 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Human Macrophages)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Human Macrophages
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Username

Dr. Francesco Elia Marino

Verified customer

Submitted Apr 06 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH=6
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
10% formalin
Username

Abcam user community

Verified customer

Submitted Feb 18 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer 1X
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 14 2015

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (peritoneal macrophages)
Permeabilization
No
Specification
peritoneal macrophages
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Varkha Agrawal

Verified customer

Submitted Jun 05 2015

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