Clone:
C9B7W
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
rat IgG1κ, rat IgG1
Applications:
FC, FA
Alternative names:
LAG-3, Ly66

Extended validation for CD223 Antibody, anti-mouse

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 C9B7W
631501-
REA776++
Cells were incubated with an excess of purified unconjugated CD223 (C9B7W) 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD223 antibodies and with a suitable counterstaining. As a control, CD223 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 CD223 (C9B7W). 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 CD223 (C9B7W). 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 CD223 (C9B7W). 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 CD223 Antibody, anti-mouse

Overview

Clone C9B7W recognizes an epitope within the D2 domain of murine CD233, also known as LAG-3, which is a 70 kDa high affinity MHC class II ligand. It is expressed on activated CD4
+
and CD8
+
α/β T cells and on a subset of NK cells. It has been reported that in association with the CD3 complex CD223 is a negative regulatory molecule to inhibit TCR-mediated signaling and to regulate homeostatic T cell expansion.

Alternative names

LAG-3, Ly66

Detailed product information

Technical specifications

CloneC9B7W
Clonalitymonoclonal
Isotyperat IgG1, rat IgG1κ
Isotype controlIsotype Control Antibody, rat IgG1
Hostrat
Type of antibodyPrimary antibodies, Functional-grade antibodies
Speciesmouse
AntigenCD223
Alternative names of antigenLAG-3, Ly66
Molecular mass of antigen [kDa]55
Distribution of antigenNK cells, T cells
Entrez Gene ID16768
RRIDAB_2656405, AB_2656422, AB_2656423, AB_2656424, AB_2752030

References for CD223 Antibody, anti-mouse

Publications

  1. Workman, C. J. et al. (2002) Cutting edge: Molecular analysis of the negative regulatory function of lymphocyte activation gene-3. J. Immunol. 169(10): 5392-5395
  2. Baixeras, E. et al. (1992) Characterization of the lymphocyte activation gene 3-encoded protein. A new ligand for human leukocyte antigen class II antigens. J. Exp. Med. 176(2): 327-337
  3. Workman, C. J. et al. (2002) Phenotypic analysis of the murine CD4-related glycoprotein, CD223 (LAG-3). Eur. J. Immunol. 32(8): 2255-2263

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