Product nameAnti-F4/80 antibody [CI:A3-1]
See all F4/80 primary antibodies
DescriptionRat monoclonal [CI:A3-1] to F4/80
SpecificityThis antibody recognises the mouse F4/80 antigen, a 160kD glycoprotein expressed by murine macrophages. Abcam does not recommend this product for IHC-P.
Tested applicationsSuitable for: WB, IP, RIA, IHC-Fr, Flow Cyt, ICC, IHC-FoFr, ICC/IF, IHC-Rmore details
Species reactivityReacts with: Mouse
Thioglycollate stimulated peritoneal macrophages from C57/BL mice
- 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.
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.
Storage instructionsShipped 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 bufferpH: 7.60
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: 0.164% Sodium phosphate, 1.45% Sodium chloride, 1% BSA
Concentration information loading...
PurityProtein G purified
Purification notesPurified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Fix & Perm / Cell Fixation & Permeabilization Kit (Flow Cytometry) (ab185917)
- Anti-TMEM119 antibody [28-3] - Microglial marker (ab209064)
- Anti-CD68 antibody [FA-11] (ab53444)
- Mouse F4/80 peptide (ab74382)
- Anti-CD11b antibody [M1/70] (ab8878)
- Anti-F4/80 antibody [F4/80] (ab90247)
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.
|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.|
FunctionOrphan 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 specificityExpression is restricted to eosinophils.
Sequence similaritiesBelongs 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 localizationCell membrane.
- Information by UniProt
- ADGRE1 antibody
- Adhesion G protein coupled receptor E1 antibody
- Adhesion G protein-coupled receptor E1 antibody
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.
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.
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?
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.
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.
This product has been referenced in:
- Xu L et al. Ablation of PPAR? in subcutaneous fat exacerbates age-associated obesity and metabolic decline. Aging Cell 17:N/A (2018). IHC-P ; Mouse . Read more (PubMed: 29383825) »
- Li D et al. CD24-p53 axis suppresses diethylnitrosamine-induced hepatocellular carcinogenesis by sustaining intrahepatic macrophages. Cell Discov 4:6 (2018). Read more (PubMed: 29423273) »