Clone:
REA126
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
ADGRE1, DD7A5-7, EGF-TM7, EMR1, Gpf480, Ly71, TM7LN3

Extended validation for F4/80 Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA126
BM8++
CI:A3-1-
T45-2342-
Cells were incubated with an excess of purified unconjugated F4/80 (REA126) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for F4/80. Peritoneal macrophages from C57BL/6 mice were stained with F4/80 antibodies and with a suitable counterstaining. As a control, F4/80 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using F4/80 (REA126). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using F4/80 (REA126). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using F4/80 (REA126). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for F4/80 Antibody, anti-mouse, REAfinity™

Overview

Clone REA126 recognizes F4/80, a member of epidermal growth factor (EGF)-transmembrane 7 (TM7) family. F4/80 is considered as one of the most specific cell-surface markers for murine macrophages. F4/80 is a seven-span transmembrane molecule with a large extracellular, multiple EGF module–containing domain. Constitutive and high expression of F4/80 is found on most resident tissue macrophages, including the spleen, microglia in the brain, Kupffer's cells in the liver, and Langerhans cells in the skin. In addition, the expression of F4/80 can also be regulated depending on the physiological status of the cell. F4/80 molecule is required for the differentiation of antigen-specific CD8
+
Tregs and is involved in inducing peripheral immune tolerance. No specific ligand for F4/80 has been reported so far.
Additional information: Clone REA126 displays negligible binding to Fc receptors.

Alternative names

ADGRE1, DD7A5-7, EGF-TM7, EMR1, Gpf480, Ly71, TM7LN3

Detailed product information

Technical specifications

CloneREA126
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenF4/80
Alternative names of antigenADGRE1, DD7A5-7, EGF-TM7, EMR1, Gpf480, Ly71, TM7LN3
Molecular mass of antigen [kDa]99
Distribution of antigenmacrophages
Entrez Gene ID13733
RRIDAB_2727574, AB_2733417, AB_2733418, AB_2733260, AB_2733261, AB_2751520, AB_2751483, AB_2784335, AB_2819539, AB_2727970

Resources for F4/80 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this
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References for F4/80 Antibody, anti-mouse, REAfinity™

Publications

  1. Lin, H-H. et al. (2005) The macrophage F4/80 receptor is required for the induction of antigen-specific efferent regulatory T cells in peripheral tolerance. J. Exp. Med. 201: 1615-1625
  2. Gordon, S. et al. (2011) F4/80 and the related adhesion-GPCRs. Eur. J. Immunol. 41(9): 2472-2476
  3. Kortlever, R. et al. (2017) Myc Cooperates with Ras by Programming Inflammation and Immune Suppression. Cell 171(6)
  4. Hume, D. A. et al. (1984) The mononuclear phagocyte system of the mouse defined by immunohistochemical localization of antigen F4/80: macrophages of bone and associated connective tissue. J. Cell. Sci. 66: 189-194
  5. Gu, C. et al. (2018) EcoHIV infection of mice establishes latent viral reservoirs in T cells and active viral reservoirs in macrophages that are sufficient for induction of neurocognitive impairment. PLoS Pathog. 14(6)
  6. Siegmund, D. et al. (2016) Activation of TNFR2 sensitizes macrophages for TNFR1-mediated necroptosis. Cell Death Dis 7(9): e2375
  7. Siegmund, D. et al. (2018) TNFR2 unlocks a RIPK1 kinase activity-dependent mode of proinflammatory TNFR1 signaling. Cell Death Dis 9(9)
  8. Lee, H. et al. (2017) Kinetics of corneal antigen presenting cells in experimental dry eye disease. BMJ Open Ophthalmol 1(1)
  9. Jia, J. et al. (2018) Cholesterol metabolism promotes B-cell positioning during immune pathogenesis of chronic obstructive pulmonary disease. EMBO Mol. Med. 10(5)

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