Clone:
53-6.7
Type of antibody:
Primary antibodies
Isotype:
rat IgG2aκ
Applications:
FC, MICS, IF, IHC
Alternative names:
Ly-2, Ly-35, Ly-B, Lyt-2, T8, CD8, Leu-2

Extended validation for CD8a 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 53-6.7
REA601++
Cells were incubated with an excess of purified unconjugated CD8a (53-6.7) 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 CD8a. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8a antibodies and with a suitable counterstaining. As a control, CD8a 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 CD8a. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8a antibodies and with a suitable counterstaining. As a control, CD8a 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 CD8a. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8a antibodies and with a suitable counterstaining. As a control, CD8a 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 CD8a. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8a antibodies and with a suitable counterstaining. As a control, CD8a 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 CD8a. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8a antibodies and with a suitable counterstaining. As a control, CD8a 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 CD8a (53-6.7). 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 CD8a (53-6.7). 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 CD8a (53-6.7). 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 CD8a Antibody, anti-mouse

Overview

Clone 53-6.7 is specific for the mouse CD8a antigen, also known as Ly-2, which is expressed on cytotoxic T cells and at lower levels on subpopulations of dendritic cells and TCRγ/δ
+
cells. It is further detected on most thymocytes (CD4
+
CD8a
+
and CD4
CD8
+
thymocytes). Cytotoxic T cells and thymocytes express CD8a as heterodimer with CD8b, whereas dendritic cells and TCRγ/δ
+
cells express a CD8a/CD8a homodimer.

Alternative names

Ly-2, Ly-35, Ly-B, Lyt-2, T8, CD8, Leu-2

Detailed product information

Technical specifications

Clone53-6.7
Clonalitymonoclonal
Isotyperat IgG2aκ
Isotype controlIsotype Control Antibody, rat IgG2a
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenCD8a
Alternative names of antigenLy-2, Ly-35, Ly-B, Lyt-2, T8, CD8, Leu-2
Molecular mass of antigen [kDa]25
Distribution of antigendendritic cells, NK cells, red blood cells, T cells, thymocytes
Entrez Gene ID12525
RRIDAB_2728078, AB_2733536, AB_2733537, AB_2752021, AB_2784288, AB_2784287, AB_2801790, AB_2801783, AB_2801982, AB_2811486, AB_2811566, AB_2659884, AB_2659885, AB_2659890, AB_2659891, AB_2728080

Resources for CD8a Antibody, anti-mouse

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