Clone:
REAL104
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC
Alternative names:
CD3-epsilon, CD3e, Leu4, T-cell surface antigen T3/Leu-4 epsilon chain, T3E, IMD18, TCRE

Extended validation for CD3 Antibody, anti-human,
REAlease
®

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 recognition with REAL104
REA613++
SK7++
OKT3++
Cells were incubated with an excess of purified unconjugated CD3 (REAL104) 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and with a suitable counterstaining. As a control, CD3 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 CD3 (REAL104). 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 CD3 (REAL104). 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 CD3 (REAL104). 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 CD3 Antibody, anti-human,
REAlease
®

Overview

Clone REAL104 is an antibody fragment derived from the full CD3 antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL104 recognizes the human CD3 antigen, a single-pass type I membrane protein also known as Leu4. CD3 is present on mature human T cells, thymocytes, a subset of NK cells, and on Purkinje cells in cerebellum. It is associated with the T cell receptor (TCR) and is responsible for its signal transduction. The CD3 antigen is a complex of five invariable chains: γ, δ, ε, ζ, and η. The epitope recognized by clone REAL104 is located on the ε-chain of the CD3 complex.
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

CD3-epsilon, CD3e, Leu4, T-cell surface antigen T3/Leu-4 epsilon chain, T3E, IMD18, TCRE

Detailed product information

Technical specifications

CloneREAL104
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Specieshuman
AntigenCD3
Alternative names of antigenCD3-epsilon, CD3e, Leu4, T-cell surface antigen T3/Leu-4 epsilon chain, T3E, IMD18, TCRE
Distribution of antigenT cells, NK cells, NKT cells
RRIDAB_2751073, AB_2751335, AB_2751328, AB_2751626, AB_2751584, AB_2751303, AB_2751292, AB_2784113, AB_2784114, AB_2784115, AB_2784116, AB_2819401, AB_2819398, AB_2751096

References for CD3 Antibody, anti-human,
REAlease
®

Publications

  1. Gold, D. P. et al. (1986) Isolation of cDNA clones encoding the 20K non-glycosylated polypeptide chain of the human T-cell receptor/T3 complex. Nature 321(6068): 431-434
  2. D'Acquisto, F. et al. (2011)
    CD3
    +
    CD4
    -
    CD8
    -
    (double negative) T cells: saviours or villains of the immune response?
    Biochem. Pharmacol. 82(4): 333-340
  3. Birnbaum, M. E. et al. (2014) Molecular architecture of the αβ T cell receptor-CD3 complex. Proc. Natl. Acad. Sci. U.S.A. 111(49): 17576-17581

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