Clone:
REA963
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
SLAMF3, Ly9, Ly-9, hly9, mLY9

Extended validation for CD229 (Ly-9) 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 REA963
Hly-9.1.25++
249936++
Cells were incubated with an excess of purified unconjugated CD229 (Ly-9) (REA963) 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 CD229 (Ly-9). Human peripheral blood mononuclear cells (PBMCs) were stained with CD229 (Ly-9) antibodies and with a suitable counterstaining. As a control, CD229 (Ly-9) 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 CD229 (Ly-9). Human peripheral blood mononuclear cells (PBMCs) were stained with CD229 (Ly-9) antibodies and with a suitable counterstaining. As a control, CD229 (Ly-9) 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 CD229 (Ly-9). Human peripheral blood mononuclear cells (PBMCs) were stained with CD229 (Ly-9) antibodies and with a suitable counterstaining. As a control, CD229 (Ly-9) 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 CD229 (Ly-9). Human peripheral blood mononuclear cells (PBMCs) were stained with CD229 (Ly-9) antibodies and with a suitable counterstaining. As a control, CD229 (Ly-9) 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 CD229 (Ly-9). Human peripheral blood mononuclear cells (PBMCs) were stained with CD229 (Ly-9) antibodies and with a suitable counterstaining. As a control, CD229 (Ly-9) 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 CD229 (Ly-9) (REA963). 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 CD229 (Ly-9) (REA963). 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 CD229 (Ly-9) (REA963). 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 CD229 (Ly-9) Antibody, anti-human, REAfinity™

Overview

Clone REA963 recognizes the human CD229 antigen, a type I transmembrane glycoprotein belonging to the CD150 subfamily of leukocyte receptors of the Ig superfamily. CD229, also known as Ly-9, has four extracellular Ig domains and a cytoplasmic domain containing two copies of the tyrosine-based motif (T-I/V-Y-x-x-V/I). Via this unique motif, CD229 interacts with SH2-binding phosphatases and the adapter molecules SAP, EAT-2, and Grb2. Expression of CD229 is found on T cells, B cells, NK cells, and thymocytes. CD229 interacts homophilically via its N-terminal V-like domain and is involved in enhancing T cell activation and Tʜ2 polarization.
Additional information: Clone REA963 displays negligible binding to Fc receptors.

Alternative names

SLAMF3, Ly9, Ly-9, hly9, mLY9

Detailed product information

Technical specifications

CloneREA963
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD229 (Ly-9)
Alternative names of antigenSLAMF3, Ly9, Ly-9, hly9, mLY9
Molecular mass of antigen [kDa]67
Distribution of antigenB cells, NK cells, T cells, thymocytes
Entrez Gene ID4063
RRIDAB_2727331, AB_2727298, AB_2727332, AB_2727299, AB_2727334, AB_2727301, AB_2727333, AB_2727300, AB_2857412, AB_2857411, AB_2727330, AB_2727297

Resources for CD229 (Ly-9) Antibody, anti-human, REAfinity™

Certificates

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References for CD229 (Ly-9) Antibody, anti-human, REAfinity™

Publications

  1. Romero, X. et al. (2004) Differential expression of SAP and EAT-2-binding leukocyte cell-surface molecules CD84, CD150 (SLAM), CD229 (Ly9) and CD244 (2B4). Tissue Antigens 64(2): 132-144
  2. Romero, X. et al. (2005) CD229 (Ly9) lymphocyte cell surface receptor interacts homophilically through its N-terminal domain and relocalizes to the immunological synapse. J. Immunol. 174(11): 7033-7042
  3. Martín, M. et al. (2005) Identification of Grb2 as a novel binding partner of the signaling lymphocytic activation molecule-associated protein binding receptor CD229. J. Immunol. 174: 5977-5986

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