CD39 Antibody, anti-human, REAfinity™
Clone:
REA739
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
Entpd1, ATPDase, NTPDase-1, SPG64, EC3.6.1.5, gp80

Extended validation for CD39 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 REA739
MZ18-23C8++
A1++
TU66++
Cells were incubated with an excess of purified unconjugated CD39 (REA739) 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.
Knockout validation
To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.
WT
KO
View details
Fluorescence microscopy image of CD39 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD39-PE (REA739, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD39 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD39-PE (REA739, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD39 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD39-PE (REA739, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD39 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD39-PE, clone (REA739). 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 propidium iodide fluorescence.
Overlay histogram showing flow cytometric analysis of CD39 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD39-PE, clone (REA739). 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 propidium iodide fluorescence.

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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39. Human peripheral blood mononuclear cells (PBMCs) were stained with CD39 antibodies and with a suitable counterstaining. As a control, CD39 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 CD39 (REA739). 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 CD39 (REA739). 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 CD39 (REA739). 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 CD39 Antibody, anti-human, REAfinity™

Overview

Clone REA739 recognizes the 70–100 kDa membrane-bound human CD39 molecule, which is expressed on an effector/memory-like subset of FoxP3
+
regulatory T cells. CD39 is an ectonucleotidase and catalyzes the hydrolysis of extracellular nucleotides, for example, ATP. Together with CD73, which is an ecto-5’-nucleotidase, this can lead to the production of adenosine. High extracellular ATP concentrations indicate tissue injury and cell death and induce various pro-inflammatory responses in immune cells.
Through its enzymatic activity, CD39 can contribute to the suppressive function of regulatory T cells, for example, by eliminating extracellular ATP or by generating adenosine, which has suppressive effects on various immune cells.
Additional information: Clone REA739 displays negligible binding to Fc receptors.

Alternative names

Entpd1, ATPDase, NTPDase-1, SPG64, EC3.6.1.5, gp80

Detailed product information

Technical specifications

CloneREA739
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
AntigenCD39
Alternative names of antigenEntpd1, ATPDase, NTPDase-1, SPG64, EC3.6.1.5, gp80
Molecular mass of antigen [kDa]58
Distribution of antigenT cells
Entrez Gene ID953
RRIDAB_2657888, AB_2657889, AB_2657890, AB_2657891, AB_2657892, AB_2657893, AB_2657894, AB_2657895, AB_2657896, AB_2657897, AB_2657898, AB_2657899, AB_2657900, AB_2904987, AB_2904986, AB_2904985, AB_2904984, AB_2657887

Resources for CD39 Antibody, anti-human, REAfinity™

Documents and Protocols

Scientific posters

Multicolor immunophenotyping of human tumor cells using MACSQuant® Analyzer 16

Scientific posters

Flow cytometric assays for CAR T cell manufacturing and patient monitoring, involving specific CAR detection reagents, stabilized pre-formulated cocktails, and automated data acquisition and analysis

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD39 Antibody, anti-human, REAfinity™

Publications

  1. Maliszewski, C. R. et al. (1994) The CD39 lymphoid cell activation antigen. Molecular cloning and structural characterization. J Immunol 153(8): 3574-3583
  2. Kaczmarek, E. et al. (1996) Identification and characterization of CD39/vascular ATP diphosphohydrolase. J. Biol. Chem. 271(51): 33116-33122
  3. Borsellino, G. et al. (2007)
    Expression of ectonucleotidase CD39 by FoxP3
    +
    reg cells: hydrolysis of extracellular ATP and immune suppression.
    Blood 110: 1225-1232

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