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

Name: CD226 (DNAM-1) Antibody, anti-human, REAfinity™
Availability: InStock

CD226 (DNAM-1) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

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

Flow cytometry applications forCD226 (DNAM-1) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright R720
Vio Bright V423
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-117-639, 130-117-490

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood cells (PBMCs) were stained with CD226 (DNAM-1) antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD56 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD226 (DNAM-1) Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

FFPE

Tested:

during development

Species tested:

human

Products used:

130-117-639, 130-117-490

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD226 (DNAM-1) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-126-485

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 clones	Overlap 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.

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.

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

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

Clone	REA1040
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis ) , chimpanzee ( Pan troglodytes ) , olive baboon ( Papio anubis )
Antigen	CD226 (DNAM-1)
Alternative names of antigen	DNAM-1, PTA-1, TLiSA1
Molecular mass of antigen [kDa]	37
Distribution of antigen	monocytes, NK cells, platelets, T cells, B cells, B cell subsets, thymocytes
Entrez Gene ID	10666
RRID	AB_2727958, AB_2728010, AB_2727959, AB_2728011, AB_2727960, AB_2728012, AB_2727961, AB_2728013, AB_2727962, AB_2889514, AB_2889513, AB_2889512, AB_2922228, AB_2922183, AB_2728009

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

Certificates

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to search for Certificates of Analysis (CoA) by lot number.

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

Publications

Shibuya, A. et al. (1996) DNAM-1, a novel adhesion molecule involved in the cytolytic function of T lymphocytes. Immunity 4: 573-581
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
Kojima, H. et al. (2003) CD226 mediates platelet and megakaryocytic cell adhesion to vascular endothelial cells. J. Biol. Chem. 278: 36748-36753
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
Moretta, L. and Moretta, A. (2004) Unravelling natural killer function: Triggering and inhibitory human NK receptors. EMBO J. 23: 255-259
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

Applications

Extended validation