Clone:
REA641
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
T cell antigen receptor complex, T3, AI504783, CD3epsilon, T3E, Ctg-3, Ctg3, T3g

Extended validation for CD3 Antibody, anti-mouse, 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 REA641
145-2C11+
17A2++
500A2+
REA606+
Cells were incubated with an excess of purified unconjugated CD3 (REA641) 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. Splenocytes from BALB/c mice 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. Splenocytes from BALB/c mice 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. Splenocytes from BALB/c mice 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. Splenocytes from BALB/c mice 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. Splenocytes from BALB/c mice 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 (REA641). 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 (REA641). 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 (REA641). 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-mouse, REAfinity™

Overview

Clone REA641 recognizes the mouse CD3 complex, which is expressed on all mature T lymphocytes, NKT cells, and during the development of thymocytes. CD3, also known as T3, belongs to the Ig superfamily. It consists of CD3ε, δ, γ, and ζ chains and forms a TCR complex by associating with TCR α/β or γ/δ chains. The TCR does not possess intracellular signaling domains, uncoupling Ag recognition from T cell signaling. The TCR is instead non-covalently associated with a multisubunit signaling apparatus, consisting of the CD3εγ and CD3εδ heterodimers and the CD3ζζ homodimer, which collectively form the TCR-CD3 complex. CD3 is involved in T cell activation, signal transduction, and antigen recognition by binding the peptide/MHC antigen complex.
Additional information: Clone REA641 displays negligible binding to Fc receptors.

Alternative names

T cell antigen receptor complex, T3, AI504783, CD3epsilon, T3E, Ctg-3, Ctg3, T3g

Detailed product information

Technical specifications

CloneREA641
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD3
Alternative names of antigenT cell antigen receptor complex, T3, AI504783, CD3epsilon, T3E, Ctg-3, Ctg3, T3g
Molecular mass of antigen [kDa]37(sum of molecular weights of subunits)
Distribution of antigenT cells, thymocytes, NKT cells
Entrez Gene ID12501, 12502
RRIDAB_2727593, AB_2751851, AB_2751822, AB_2801978, AB_2751847, AB_2752207, AB_2801803, AB_2819528, AB_2727569

Resources for CD3 Antibody, anti-mouse, REAfinity™

References for CD3 Antibody, anti-mouse, REAfinity™

Publications

  1. 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
  2. Gold, D. P. et al. (1987) Evolutionary relationship between the T3 chains of the T-cell receptor complex and the immunoglobulin supergene family. Proc. Natl. Acad. Sci. U.S.A. 84(21): 7649-7653
  3. Miescher, G. C. et al. (1989) Production and characterization of a rat monoclonal antibody against the murine CD3 molecular complex. Immunol. Lett. 23(2): 113-118
  4. Exley, M. et al. (1991) Structure, assembly and intracellular transport of the T cell receptor for antigen. Semin. Immunol. 3(5): 283-297

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