CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™

Name: CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™
Availability: InStock

CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA975

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC

Alternative names:

CD3e, CD3epsilon, T3E

Intracellular flow cytometry applications forCD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™

APC
FITC
PE
Vio B515
Vio R667

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-116-290, 130-116-187

Protocols:

Intracellular flow cytometry staining protocol (Inside Stain Kit 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were fixed, permeabilizied, and stained with CD3ε (intracellular) antibodies or with the corresponding REA Control (I) antibodies (left images). Flow cytometry was performed using the MACSQuant^®Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals.

Extended validation for CD3ε (intracellular) Antibody, anti-human/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 clones	Overlap in epitope recognition with REA975
APA1/1	++
UCHT1	-
CD3-12	+
Cells were incubated with an excess of purified unconjugated CD3ε (intracellular) (REA975) 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.

Flow cytometric comparison of different clones for CD3ε (intracellular). Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with CD3ε (intracellular) antibodies. As a control, CD3ε (intracellular) 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 Viobility 405/520 Fixable Dye 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 CD3ε (intracellular). Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with CD3ε (intracellular) antibodies. As a control, CD3ε (intracellular) 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 Viobility 405/520 Fixable Dye 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 CD3ε (intracellular). Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with CD3ε (intracellular) antibodies. As a control, CD3ε (intracellular) 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 Viobility 405/520 Fixable Dye 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 CD3ε (intracellular). Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with CD3ε (intracellular) antibodies. As a control, CD3ε (intracellular) 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 Viobility 405/520 Fixable Dye 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 CD3ε (intracellular). Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with CD3ε (intracellular) antibodies. As a control, CD3ε (intracellular) 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 Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA975 recognizes an intracellular part of the CD3ε chain (activation epitope). CD3ε is a single-pass type I membrane protein, which is a part of the CD3 complex and a subunit of the TCR complex. The CD3 complex mediates signal transduction in several intracellular pathways and has a role in T cell activation and antigen recognition by binding the peptide/MHC antigen complex. CD3ε is expressed on all mature T lymphocytes, NKT cells, and during the development of thymocytes.

Additional information: Clone REA975 displays negligible binding to Fc receptors.

Alternative names

CD3e, CD3epsilon, T3E

Detailed product information

Technical specifications

Clone	REA975
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, mouse
Antigen	CD3ε (intracellular)
Alternative names of antigen	CD3e, CD3epsilon, T3E
Molecular mass of antigen [kDa]	21
Distribution of antigen	T cells, NKT cells, thymocytes
Entrez Gene ID	916
RRID	AB_2727389, AB_2727452, AB_2727390, AB_2727453, AB_2727391, AB_2727888, AB_2727851, AB_2727889, AB_2727852, AB_2727451

Resources for CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD3ε (intracellular) Antibody, anti-human/mouse, REAfinity™

Publications

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
Clevers, H. et al. (1988) Characterization and expression of the murine CD3-epsilon gene. Proc. Natl. Acad. Sci. U.S.A. 85(22): 8623-8627
Soudais, C. et al. (1993) Independent mutations of the human CD3-epsilon gene resulting in a T cell receptor/CD3 complex immunodeficiency. Nat. Genet. 3(1): 77-81

Applications

Extended validation