Clone:
REA1040
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
DNAM-1, PTA-1, TLiSA1

Extended validation for CD226 (DNAM-1) 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 REA1040
DX11++
11A8++
TX25++
Cells were incubated with an excess of purified unconjugated CD226 (DNAM-1) (REA1040) 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 CD226 (DNAM-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD226 (DNAM-1) antibodies and with a suitable counterstaining. As a control, CD226 (DNAM-1) 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 CD226 (DNAM-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD226 (DNAM-1) antibodies and with a suitable counterstaining. As a control, CD226 (DNAM-1) 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 CD226 (DNAM-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD226 (DNAM-1) antibodies and with a suitable counterstaining. As a control, CD226 (DNAM-1) 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 CD226 (DNAM-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD226 (DNAM-1) antibodies and with a suitable counterstaining. As a control, CD226 (DNAM-1) 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 CD226 (DNAM-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD226 (DNAM-1) antibodies and with a suitable counterstaining. As a control, CD226 (DNAM-1) 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 CD226 (DNAM-1) (REA1040). 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 CD226 (DNAM-1) (REA1040). 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 CD226 (DNAM-1) (REA1040). 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 CD226 (DNAM-1) Antibody, anti-human, REAfinity™

Overview

Clone REA1040 recognizes the human CD226 (DNAM-1) antigen, a 65 kDa transmembrane glycoprotein expressed on T cells, NK cells, platelets, monocytes, and a subset of B cells. It is also expressed by a subset of CD3
+
thymocytes. The surface association of CD226 (DNAM-1) with LFA-1 on NK cells and T cells permits the transduction of cytolytic signals upon binding of CD226 (DNAM-1) to its ligands CD155 (PVR) and CD112 (Nectin-2).
Additional information: Clone REA1040 displays negligible binding to Fc receptors.

Alternative names

DNAM-1, PTA-1, TLiSA1

Detailed product information

Technical specifications

CloneREA1040
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
)
,
olive baboon (
Papio anubis
)
AntigenCD226 (DNAM-1)
Alternative names of antigenDNAM-1, PTA-1, TLiSA1
Molecular mass of antigen [kDa]37
Distribution of antigenmonocytes, NK cells, platelets, T cells, B cells, B cell subsets, thymocytes
Entrez Gene ID10666
RRIDAB_2727958, AB_2728010, AB_2727959, AB_2728011, AB_2727960, AB_2728012, AB_2727961, AB_2728013, AB_2727962, AB_2728009

Resources for CD226 (DNAM-1) Antibody, anti-human, REAfinity™

Certificates

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

References for CD226 (DNAM-1) Antibody, anti-human, REAfinity™

Publications

  1. Shibuya, A. et al. (1996) DNAM-1, a novel adhesion molecule involved in the cytolytic function of T lymphocytes. Immunity 4: 573-581
  2. Bottino, C. et al. (2003) Identification of PVR (CD155) and Nectin-2 (CD112) as cell surface ligands for the human DNAM-1 (CD226) activating molecule. J. Exp. Med. 198: 557-567
  3. Kojima, H. et al. (2003) CD226 mediates platelet and megakaryocytic cell adhesion to vascular endothelial cells. J. Biol. Chem. 278: 36748-36753
  4. Castriconi, R. et al. (2004) Natural killer cell-mediated killing of freshly isolated neuroblastoma cells: critical role of DNAX accesory molecule-1-poliovirus receptor interaction. Cancer Res. 64: 9180-9184
  5. Moretta, L. and Moretta, A. (2004) Unravelling natural killer function: Triggering and inhibitory human NK receptors. EMBO J. 23: 255-259
  6. Pende, D. et al. (2005) Analysis of the receptor-ligand interactions in the natural killer-mediated lysis of freshly isolated myeloid or lymphoblastic leukemias: evidence for the involvement of the poliovirus receptor (CD155) and Nectin-2 (CD112). Blood 105: 2066-2073

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