Exbio — Research products — Antibodies — CD and related antigens — Anti-CD3 epsilon (activation epitope) Purified

Anti-CD3 epsilon (activation epitope) Purified

Regulatory status

RUO

Antigen

CD3 epsilon (activation epitope)

Clone

APA1/1

Format

Purified

Reactivity

Mouse, Human

Application

IP, WB, FC (QC tested), IHC(F), ICC

Variant

0.1 mg

11-599-C100

In stock

150.00 USD

0.025 mg

11-599-C025

Delivery 1 week

75.00 USD

Variant

0.1 mg

11-261-C100

In stock

150.00 USD

0.1 mg

11-261-C100

In stock

75.00 USD

Contact distributor

Product details

Description

Images

References

Isotype

Mouse IgG1

Specificity

The mouse monoclonal antibody APA1/1 recognizes an activation-dependent intracellular epitope of CD3 epsilon. Exposure of the epitope precedes CD3 phosphorylation and recruitment and activation of ZAP70, which initiates the signaling cascade produced by T-cell activation. APA1/1 provides the earliest known marker for TCR-mediated T cell activation.

Application

IP, WB, FC (QC tested), IHC(F), ICC

Application details

Flow cytometry: Intracellular staining; recommended dilution: 1-4 µg/ml; positive control: human T cells stimulated with anti-CD3 (MEM-57) antibody (1 μg/ml). Sample preparation: At the end of stimulation of T cells, perform staining of surface markers (if required) in PBS + 0.1% BSA for 20 min. on ice. Wash with PBS and fix with 2% formaldehyde, 30 min on ice. Wash with PBS and incubate in PBS + 0,1% saponine, 5 min. RT. Incubate the cells in PBS + 1% BSA + 0.03% saponine, 15 min. on ice. Incubate with fluorescence-labeled APA1/1 antibody (1-2 μg/ml) in PBS + 1% BSA + 0.03% saponine in dark, 20 min. RT. Wash with PBS + 1% BSA + 0.03% saponine, resuspend in PBS.
Immunocytochemistry: Fixed and permeabilised cells. The antibody can distinguish TCR-stimulated from non-stimulated cells.

Reactivity

Mouse, Human

Immunogen

Purified human CD3 proteins isolated from thymus

Concentration

1 mg/ml

Preparation

Purified by protein-A affinity chromatography.

Formulation

Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

Storage and handling

Store at 2-8°C. Do not freeze.

The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.

Other names

CD3 epsilon chain, T3E, IMD18

Antigen description

CD3 complex is crucial in transducing antigen-recognition signals into the cytoplasm of T cells and in regulating the cell surface expression of the TCR complex. T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta. These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR-induced growth arrest, cell survival and proliferation.

Entrez Gene ID 916

UniProt ID P07766

Separation of Jurkat cells stained anti-human CD3 activation epitope (APA1/1) purified antibody (concentration in sample 5 μg/ml, GAM FITC, red-filled) from Jurkat cells unstained by primary antibody (GAM FITC, black-dashed) in flow cytometry analysis (intracellular staining).

Product specific references:

Risueño RM, van Santen HM, Alarcón B: A conformational change senses the strength of T cell receptor-ligand interaction during thymic selection. Proc Natl Acad Sci U S A. 2006 Jun 20;103(25):9625-30.
PubMed

Alarcón B, Ley SC, Sánchez-Madrid F, Blumberg RS, Ju ST, Fresno M, Terhorst C: The CD3-gamma and CD3-delta subunits of the T cell antigen receptor can be expressed within distinct functional TCR/CD3 complexes. EMBO J. 1991 Apr;10(4):903-12.
PubMed

Gil D, Schrum AG, Alarcón B, Palmer E: T cell receptor engagement by peptide-MHC ligands induces a conformational change in the CD3 complex of thymocytes. J Exp Med. 2005 Feb 21;201(4):517-22.
PubMed

Risueño RM, Gil D, Fernández E, Sánchez-Madrid F, Alarcón B: Ligand-induced conformational change in the T-cell receptor associated with productive immune synapses. Blood. 2005 Jul 15;106(2):601-8.
PubMed

Delgado P, Fernández E, Dave V, Kappes D, Alarcón B: CD3delta couples T-cell receptor signalling to ERK activation and thymocyte positive selection. Nature. 2000 Jul 27;406(6794):426-30.
PubMed

Tailor P, Tsai S, Shameli A, Serra P, Wang J, Robbins S, Nagata M, Szymczak-Workman AL, Vignali DA, Santamaria P: The proline-rich sequence of CD3epsilon as an amplifier of low-avidity TCR signaling. J Immunol. 2008 Jul 1;181(1):243-55.
PubMed

Rieux-Laucat F, Hivroz C, Lim A, Mateo V, Pellier I, Selz F, Fischer A, Le Deist F: Inherited and somatic CD3zeta mutations in a patient with T-cell deficiency. N Engl J Med. 2006 May 4;354(18):1913-21.
PubMed

Torres PS, Alcover A, Zapata DA, Arnaud J, Pacheco A, Martín-Fernández JM, Villasevil EM, Sanal O, Regueiro JR: TCR dynamics in human mature T lymphocytes lacking CD3 gamma. J Immunol. 2003 Jun 15;170(12):5947-55.
PubMed

Wong AP, Dutly AE, Sacher A, Lee H, Hwang DM, Liu M, Keshavjee S, Hu J, Waddell TK: Targeted cell replacement with bone marrow cells for airway epithelial regeneration. Am J Physiol Lung Cell Mol Physiol. 2007 Sep;293(3):L740-52.
PubMed

Variant

0.1 mg

11-599-C100

In stock

150.00 USD

0.025 mg

11-599-C025

Delivery 1 week

75.00 USD

Variant

0.1 mg

11-261-C100