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Safety Data Sheet (SDS) Purified Anti-human/mouse CD3epsilon Activation Epitope Antibody     Product Data Sheet (PDF)    
Purified Anti-human/mouse CD3ε (activation Epitope) Antibody
2413505 50 µg $50.00       
2413510 500 µg $225.00       
Clone: APA1/1
Isotype: Mouse IgG1, κ
Reactivity: Human, Mouse
Immunogen: Purified denatured CD3 proteins from human thymus
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application:

ICFC - Quality tested
WB - Validated
IF, in vivo cell tracking - Reported in the literature
ICC, IHC, IP - Reported by the developer

Application Notes:

Clone APA1/1 recognizes an activation-dependent intracellular epitope of the CD3ε chain. Additional reported applications (for the relevant formats of this clone) include: immunofluorescent staining of paraformaldehyde fixed sections1, in vivo tracking of T cells 2, and Western blotting3. The LEAF purified format is recommended for in vivo studies. 

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. For Western blotting, the suggested use of this reagent is 1.0 - 4.0 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application References:

1. Salmerón A, et al. 1991. J. Immunol. 147:3047. (ICFC, IF)
2. Risueño RM, et al. 2005. Blood. 106:601. (IF, in vivo tracking, Mouse Reactivity)
3. Risueño RM, et al. 2008. PLos One. 3:e1747. (WB)
4. Gil D, et al. 2005. J. Exp. Med. 201:517.
5. Risueño RM, et al. 2006. Proc. Natl. Acad. Sci. USA. 103:9625.

Human peripheral blood lymphocytes were

Human peripheral blood lymphocytes were fixed and permeabilized with 70% ethanol, followed by intracellular staining with purified CD3e (clone APA1/1, filled histogram) or mouse IgG1, κ isotype control (open histogram) followed by anti-mouse IgG PE.

Western blot analysis of extracts

Western blot analysis of extracts from Jurkat (lane 1) and PBMC untreated (lane 2) and PBMC activated with CD3/CD28(lane 3) cells using anti-CD3ε (activation epitope) antibody (clone APA1/1). β-actin (poly6221) was used as loading control.



Description:

The CD3ε chain of the CD3-TCR complex is a type I membrane protein that undergoes conformational change following ligation of the TCR-CD3 complex.  This conformational change leads to exposure of the proline rich sequence in CD3ε and results in recruitment of Nck. It is expressed by all mature T cells and forms a heteromeric complex with CD3 α/β or γ/δ and ζ chains. It is important in T cell development, and deficiency of this gene has been associated with severe immunodeficiency.

Other Names: CD3e molecule, epsilon (CD3-TCR complex), CD3e antigen, CD3 epsilon
Structure: Type I membrane protein, part of the TCR-CD3 complex. 207 amino acids, predicted molecular weight of 23 kD.
Distribution: Mature T cells.
Function: Important role in T cell development and in linking antigen recognition with intracellular signaling pathways.
Ligand Receptor: MHC antigen.
Interaction: Interacts with Nck.
Antigen References:

1. Savage PD, et al. 1988. Cytogenet. Cell. Genet. 49:289.
2. DeJarnette JB, et al. 1998. Proc. Natl. Acad. Sci USA. 95:14909.
3. Wang B, et al. 1999. J. Immunol. 162:88.
4. Szymczak AL, et al. 2005. J. Immunol. 175:270.
5. Borroto A, et al. 2014. J. Immunol. 192:2042.