Clone:
REA804
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
NT5E, CALJA, E5NT, NT, Nt5, NTE, eN, eNT

Extended validation for CD73 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 REA804
AD2++
Cells were incubated with an excess of purified unconjugated CD73 (REA804) 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 CD73. Human peripheral blood mononuclear cells (PBMCs) were stained with CD73 antibodies and with a suitable counterstaining. As a control, CD73 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 CD73. Human peripheral blood mononuclear cells (PBMCs) were stained with CD73 antibodies and with a suitable counterstaining. As a control, CD73 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 CD73. Human peripheral blood mononuclear cells (PBMCs) were stained with CD73 antibodies and with a suitable counterstaining. As a control, CD73 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 CD73. Human peripheral blood mononuclear cells (PBMCs) were stained with CD73 antibodies and with a suitable counterstaining. As a control, CD73 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 CD73 (REA804). 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 CD73 (REA804). 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 CD73 (REA804). 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 CD73 Antibody, anti-human, REAfinity™

Overview

Clone REA804 recognizes the human CD73 antigen, which is a 70 kDa membrane-bound enzyme that catalyzes the conversion of AMP to bioactive adenosine at neutral pH. This glycoprotein is also called ecto-5’-nucleotidase. CD73 is present on subsets of T and B lymphocytes, follicular dendritic cells, endothelial cells, and marrow stromal cells (MSCs).
Additional information: Clone REA804 displays negligible binding to Fc receptors.

Alternative names

NT5E, CALJA, E5NT, NT, Nt5, NTE, eN, eNT

Detailed product information

Technical specifications

CloneREA804
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
)
,
chimpanzee (
Pan troglodytes
)
AntigenCD73
Alternative names of antigenNT5E, CALJA, E5NT, NT, Nt5, NTE, eN, eNT
Distribution of antigenB cells, dendritic cells, endothelial cells, epithelial cells, lymphocytes, stem cells, T cells, mesenchymal stem cells
Entrez Gene ID4907
RRIDAB_2659165, AB_2659166, AB_2659167, AB_2659168, AB_2659169, AB_2659170, AB_2659171, AB_2784276, AB_2784275, AB_2659172, AB_2659173, AB_2659164

References for CD73 Antibody, anti-human, REAfinity™

Publications

  1. Thomson, L. F. et al. (1990) Production and characterization of monoclonal antibodies to the glycosyl Tissue Antigens 35: 9-19
  2. Hansen, K. R. et al. (1995) Isolation and characterization of the promoter of the human 5'-nucleotidase (CD73)-encoding gene. Gene 167(1-2): 307-312
  3. Pittenger, M. F. et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284: 143-147

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