Clone:
REA978
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
FCGR1A, CD64A, FcR I, IGFR1, FcγRI

Extended validation for CD64 Antibody, anti-human, 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 REA978
10.1++
10.1.1++
Cells were incubated with an excess of purified unconjugated CD64 (REA978) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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 (REA978). 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 (REA978). 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 (REA978). 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-human, REAfinity™

Overview

Clone REA978 recognizes the human CD64 antigen, a high affinity Fc receptor for IgG, also known as FcγRI. CD64 contains three Ig-like domains in its extracellular domain and binds both monomeric and aggregated IgG and contributes to antibody-dependent cellular cytotoxicity (ADCC), phagocytosis, and the clearance of immune complexes. It is constitutively expressed on macrophages, monocytes, and eosinophils and can be up-regulated on neutrophils during infection or after stimulation with IFN-γ and colony-stimulating factors G-CSF and GM-CSF.
Additional information: Clone REA978 displays negligible binding to Fc receptors.

Alternative names

FCGR1A, CD64A, FcR I, IGFR1, FcγRI

Detailed product information

Technical specifications

CloneREA978
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
,
chimpanzee (
Pan troglodytes
)
, baboon, capuchin monkey,
squirrel monkey (
Saimiri sciureus
)
AntigenCD64
Alternative names of antigenFCGR1A, CD64A, FcR I, IGFR1, FcγRI
Molecular mass of antigen [kDa]41
Distribution of antigengranulocytes, macrophages, monocytes, eosinophils, neutrophils
Entrez Gene ID2209
RRIDAB_2727393, AB_2727456, AB_2727394, AB_2727457, AB_2727395, AB_2727461, AB_2727399, AB_2727462, AB_2727400, AB_2727463, AB_2727401, AB_2727458, AB_2727396, AB_2727459, AB_2727397, AB_2727460, AB_2727398, AB_2727454, AB_2727392, AB_2811651, AB_2727455

References for CD64 Antibody, anti-human, REAfinity™

Publications

  1. Shen, L. et al. (1987) Polymorphonuclear leukocyte function triggered through the high affinity Fc receptor for monomeric IgG. J. Immunol. 139(2): 534-538
  2. Buckle, A. M. and Hogg, N. (1989) The effect of IFN-gamma and colony-stimulating factors on the expression of neutrophil cell membrane receptors. J. Immunol. 143(7): 2295-2301
  3. Herra, C. et al. (1996) Increased expression of Fc gamma receptors on neutrophils and monocytes may reflect ongoing bacterial infection. J. Med. Microbiol. 44(2): 135-140

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