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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
Out of stock
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
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
Other names
CD3 epsilon chain, T3E, IMD18
Concentration
1 mg/ml
Preparation
Purified by protein-A affinity chromatography
Formulation
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Storage and handling
Store at 2-8°C. Do not freeze. Do not use after expiration date stamped on vial label.
Exbio licence note
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.
Flow cytometry intracellular staining pattern of human peripheral blood stained using anti-CD3 epsilon activation epitope (APA1/1) purified antibody (concentration in sample 4 μg/ml, GAM FITC, red) and human peripheral blood stained using Mouse IgG1 Isotype Control (MOPC-21) purified antibody (concentration in sample 4 μg/ml, GAM FITC, blue).
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
Out of stock
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
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