Clone:
REA734
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
Leu-2, CD8a, MAL, p32, Cd8b1, LY3, Lyt3, P37, Ly-2

Extended validation for CD8 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 REA734
37006++
OKT8++
BW135/80+
HIT8a++
REAL100++
RPA-T8-
SK1++
Cells were incubated with an excess of purified unconjugated CD8 (REA734) 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 (REA734). 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 (REA734). 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 (REA734). 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, REAfinity™

Overview

Clone REA734 recognizes the human CD8 antigen, a cell-surface glycoprotein also known as Leu-2 or CD8a. CD8 is strongly expressed on human cytotoxic T cells and thymocytes as well as on a subset of natural killer (NK) cells. The CD8 antigen is a disulfide-linked dimer that exists either as a CD8a homodimer or as a CD8α/β heterodimer. CD8 acts as a coreceptor for the T cell receptor and binds to the MHC class I molecules. CD8 is involved in T cell development and activation of mature T cells. The CD8 antibody recognizes the α-subunit of the antigen.
Additional information: Clone REA734 displays negligible binding to Fc receptors.

Alternative names

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

Detailed product information

Technical specifications

CloneREA734
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD8
Alternative names of antigenLeu-2, CD8a, MAL, p32, Cd8b1, LY3, Lyt3, P37, Ly-2
Molecular mass of antigen [kDa]45
Distribution of antigenNK cells, T cells
Entrez Gene ID925, 926
RRIDAB_2659233, AB_2659234, AB_2659235, AB_2659236, AB_2659237, AB_2659238, AB_2659239, AB_2659240, AB_2659241, AB_2659242, AB_2659243, AB_2659244, AB_2659245, AB_2659246, AB_2659247, AB_2659248, AB_2659249, AB_2751050, AB_2751048, AB_2659250, AB_2659251, AB_2801862, AB_2659232

Reviews for CD8 Antibody, anti-human, REAfinity™

Human Anti-CD8-APC Antibody

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CD8-APC, human (130-110-679)

This anti-CD8 antibody was tested in order to establish a panel to describe and distinguish peripheral blood lymphocytes (from human PBMCs). Clone REA734 recognizes the human CD8 antigen, a cell-surface glycoprotein also known as Leu-2 or CD8a. We are particularly interested in the innate compartment of lymphocytes and we use CD8 to target, and also exclude, a particular subset of T lymphocytes.

References for CD8 Antibody, anti-human, REAfinity™

Publications

  1. Devine, L. et al. (1999) Orientation of the Ig domains of CD8 alpha beta relative to MHC class I. J. Immunol. 162(2): 846-851
  2. Gao, G. F. et al. (2000) Molecular interactions of coreceptor CD8 and MHC class I: the molecular basis for functional coordination with the T-cell receptor. Immunol. Today 21(12): 630-636

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