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
  • Host species
    Rat
  • 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
    • WB: Mouse spleen and macrophages lysate. ICC/IF: Raw264.7 cells; Mouse bone marrow cells. IHC-Fr: Mouse spleen and intestine tissue. Flow Cyt: J774 cells and peritoneal macrophages.
  • 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
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • 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.

We recommend ab150165 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) secondary antibody.

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

Images

  • 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.

  • 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

  • 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
    All lanes : 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?

    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

This product has been referenced in:
  • Greenhalgh SN  et al. Loss of Integrin avß8 in Murine Hepatocytes Accelerates Liver Regeneration. Am J Pathol 189:258-271 (2019). Read more (PubMed: 30448409) »
  • Liu B  et al. Intermittent Fasting Improves Glucose Tolerance and Promotes Adipose Tissue Remodeling in Male Mice Fed a High-Fat Diet. Endocrinology 160:169-180 (2019). Read more (PubMed: 30476012) »
See all 398 Publications for this product

Customer reviews and Q&As

81-86 of 86 Abreviews or Q&A

Answer


Ab6640 binds to macrophages from different sites including the peritoneal cavity, lung, spleen and thymus, to blood monocytes and to macrophages derived from bone marrow precursors in culture. The antigen is not expressed by lymphocytes or polymorphonuclear cells.
CD11b is implicated in various adhesive interactions of monocytes, macrophages and granulocytes as well as in mediating the uptake of complement coated particles. It is identical with CR3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the RGD peptide in C3b. CD11b is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain. The Mac1 CD11b antigen is present on macrophages, granulocytes, natural killer cells, blood monocytes. CD11b is expressed on 8% spleen cells, 44% bone marrow cells and less than 1% of thymocytes.
Ab6640 recognizes the mouse F4/80 antigen, a 160kD glycoprotein expressed by murine macrophages. I don't know which species you are working on, but this antibody has been tested for cross-reactivity with mouse. It has also been tested for application in IHC.

Read More

Answer

Ab6640 binds to macrophages from different sites including the peritoneal cavity, lung, spleen and thymus, to blood monocytes and to macrophages derived from bone marrow precursors in culture. The antigen is not expressed by lymphocytes or polymorphonuclear cells. The antibody does react with murine Kupffer cells.

Read More

Answer

The originator of this antibody was able to provide the following information. "We would normally expect to see cell membrane staining. Unfortunately, I do not have any pictures. Although, obviously as looking at whole cells there will be brown staining across the cell when looking at them microscopically."

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Answer

This antibody binds to macrophages from different sites including the peritoneal cavity, lung, spleen and thymus, to blood monocytes and to macrophages derived from bone marrow precursors in culture. The immunogen used to produce this ab6640 were thioglycollate stimulated peritoneal macrophages from C57/BL mice. So yes, it does seem that ab6640 may be a good choice for you.
We don't have a specific IHC protocol but would recommend referring to our general IHC protocol which can be found in the protocols tab on the online datasheet. Please note that this antigen is known to be sensitive to fixation: for less than 1 day fixation either no antigen retrieval, Trypsin or Proteinase K (6 min, RT) enzyme antigen retrieval is recommended.

Read More

Answer

We are pleased to confirm that the anti-mouse F4/80 antibody does react with murine Kupffer cells.

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Question
Answer

This antibody is suitable for use in brain, where it stains microglia.

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81-86 of 86 Abreviews or Q&A

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