Clone:
REA286
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
FCGR1, FcgammaRI, IGGHAFC, FcR I, FcγRI

Extended validation for CD64 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 REA286
X54-5/7.1++
Cells were incubated with an excess of purified unconjugated CD64 (REA286) 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 CD64. Bone marrow cells from C57BL/6 mice were stained with CD64 antibodies and with a suitable counterstaining. As a control, CD64 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 CD64. Bone marrow cells from C57BL/6 mice were stained with CD64 antibodies and with a suitable counterstaining. As a control, CD64 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 CD64. Bone marrow cells from C57BL/6 mice were stained with CD64 antibodies and with a suitable counterstaining. As a control, CD64 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 CD64. Bone marrow cells from C57BL/6 mice were stained with CD64 antibodies and with a suitable counterstaining. As a control, CD64 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 CD64 (REA286). 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 CD64 (REA286). 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 CD64 (REA286). 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 CD64 Antibody, anti-mouse, REAfinity™

Overview

Clone REA286 recognizes the CD64 antigen, a 72 kDa integral membrane glycoprotein also known as high affinity immunoglobulin gamma Fc receptor I (FcγRI). Structurally CD64 is composed of a signal peptide that allows its transport to the surface of a cell, three extracellular immunoglobulin domains of the C2-type that it uses to bind antibody, a hydrophobic transmembrane domain, and a short cytoplasmic tail. CD64 is constitutively expressed on macrophages, monocytes, dendritic cells, and mast cells. Treatment with cytokines like IFN-γ and G-CSF induces CD64 upregulation. CD64 binds monomeric IgG-type antibodies with high affinity and mediates endocytosis, phagocytosis, antibody-dependent cellular toxicity, cytokine release, and superoxide generation.
Additional information: Clone REA286 displays negligible binding to Fc receptors.

Alternative names

FCGR1, FcgammaRI, IGGHAFC, FcR I, FcγRI

Detailed product information

Technical specifications

CloneREA286
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD64
Alternative names of antigenFCGR1, FcgammaRI, IGGHAFC, FcR I, FcγRI
Molecular mass of antigen [kDa]42
Distribution of antigendendritic cells, macrophages, monocytes, mast cells
Entrez Gene ID14129
RRIDAB_2751786, AB_2751727, AB_2751554, AB_2751553

References for CD64 Antibody, anti-mouse, REAfinity™

Publications

  1. Tan, P. S. et al. (2003) Unique monoclonal antibodies define expression of FcgRI on macrophages and mast cell lines and demonstrate heterogeneity among subcutaneous and other dendritic cells. J. Immunol. 170: 2549-2556
  2. Ioan-Facsinay, A. et al. (2002) FcgammaRI (CD64) contributes substantially to severity of arthritis, hypersensitivity responses, and protection from bacterial infection. Immunity 16(3): 391-402
  3. Nimmerjahn, F. et al. (2006) Fcgamma receptors: old friends and new family members. Immunity 24(1): 19-28

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