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.

PRODUCT DETAILS

Clone: APA1/1

Specificity: CD3 Epsilon (Activation Epitope)

Reactivity: Human|Mouse

Format: Purified

Isotype: Mouse IgG1

Application: IP, WB, ICC, FC-IC

Storage: 2-8°C protected from light.Do not freeze

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

Data Sheets: TDS | SDS

RECOMMENDED USAGE

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.

REFERENCES

1. Risueño RM, et al. 2006. Proc Natl Acad Sci U S A 103:9625-30.
2. Alarcón B, et al. 1991. EMBO J 10:903-12.
3. Gil D, et al. 2005. J Exp Med 201:517-22.
4. Risueño RM, et al. 2005. Blood 106:601-8.
5. Delgado P, et al. 2000. Nature 406:426-30.
6. Tailor P, et al. 2008. J Immunol 181:243-55.
7. Rieux-Laucat F, et al. 2006. N Engl J Med 354:1913-21.
8. Torres PS, et al. 2003. J Immunol 170:5947-55.
9. Wong AP, et al. 2007. Am J Physiol Lung Cell Mol Physiol 293:L740-52.

For research use only. Not intended for use in diagnostic procedures.