Clone:
BW135/80
Type of antibody:
Primary antibodies
Isotype:
mouse IgG2aκ
Applications:
FC, MICS, IF, IHC, ICC
Alternative names:
Ly-2, Leu-2, CD8a, MAL, p32, Cd8b1, LY3, Lyt3, P37

Extended validation for CD8 Antibody, anti-human

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 BW135/80
37006+
OKT8+
REA734+
HIT8a+
REAL100+
RPA-T8++
SK1+
Cells were incubated with an excess of purified unconjugated CD8 (BW135/80) 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8. Human peripheral blood mononuclear cells (PBMCs) were stained with CD8 antibodies and with a suitable counterstaining. As a control, CD8 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 CD8 (BW135/80). 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 CD8 (BW135/80). 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 CD8 (BW135/80). 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 CD8 Antibody, anti-human

Overview

The monoclonal antibody BW135/80 recognizes the human CD8 antigen which is strongly expressed on human cytotoxic T cells and thymocytes, and is also expressed on a subset of NK cells. The CD8 antigen is a disulfide-linked dimer that exists either as a CD8α homodimer or as a CD8α/β heterodimer. CD8 acts as a co-receptor for the T cell receptor and binds to the MHC class I molecule. The CD8 antibody recognizes the α-subunit of the antigen.

Alternative names

Ly-2, Leu-2, CD8a, MAL, p32, Cd8b1, LY3, Lyt3, P37

Detailed product information

Technical specifications

CloneBW135/80
Clonalitymonoclonal
Isotypemouse IgG2aκ
Isotype controlIsotype Control Antibody, mouse IgG2a
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
AntigenCD8
Alternative names of antigenLy-2, Leu-2, CD8a, MAL, p32, Cd8b1, LY3, Lyt3, P37
Molecular mass of antigen [kDa]45(sum of molecular weights of subunits)
Distribution of antigendendritic cells, NK cells, red blood cells, T cells, thymocytes
Entrez Gene ID925
RRIDAB_2725985, AB_2726266, AB_2725991, AB_2726261, AB_2725986, AB_2726257, AB_2725982, AB_2726265, AB_2725990, AB_2726267, AB_2725992, AB_2726263, AB_2725988, AB_2726262, AB_2725987, AB_2726258, AB_2725983, AB_2726264, AB_2725989, AB_2726259, AB_2725984, AB_2726260

References for CD8 Antibody, anti-human

Publications

  1. Jonker, M. et al. (1989) Reactivity of mAb specific for human CD markers with rhesus monkey leucocytes. Oxford, Oxford University Press (Leukocyte Typing IV): 1058-1063

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