CD1d is a membrane protein non-covalently bonded to β2-microglobulin (β2m) and shows high homology between human and mouse. CD1d can present α-galactosylceramide (α-GalCer), a glycolipid extracted and isolated from the marine sponge, and this complex can activate human and murine CD1d-restricted NKT cells. CD1d tetramer is a reagent prepared by tetramerization of complexes of CD1d and β2m by PE- or APC- labeled streptavidin. Binding this reagent to α-GalCer enables highly sensitive detection of CD1d-restricted NKT cells and can be combined with antibodies to study NKT cell function by flow cytometry.
Dissolving and loading α-GalCer on CD1d tetramer
| Product | a-GalCer unloaded Code no. | a-GalCer loaded Code no. |
|---|---|---|
| Human CD1d Tetramer-PE | TS-HCD-1 | TS-HCG-1 |
| Human CD1d Tetramer-APC | TS-HCD-2 | TS-HCG-2 |
| Mouse CD1d Tetramer-PE | TS-MCD-1 | TS-MCG-1 |
| Mouse CD1d Tetramer-APC | TS-MCD-2 | TS-MCG-2 |
Detection of human NKT cells with CD1d tetramer
PBMC were separated from peripheral blood of healthy subjects and incubated at room temperature for 5 minutes with 40 ul
of Clear Back (Human Fc receptor blocking reagent, MBL code no. MTG-001). CD3-FITC and human CD1d tetramer-PE (with or without binding of α-GalCer) were added and incubated for 30 minutes at 4°C protected from light, and cells were analyzed by flow cytometry. Results showed total cells contained 0.05% NKT cells, as defined by CD1d/CD3 dual positivity.
References
- Stephane S, et al., J. Immunol. Methods 268: 107-121 (2002)
- Matsuda JL, et al., J. Exp. Med. 192: 741-754 (2000)
- Benlagha K, et al., J. Exp. Med. 191: 1895-1903 (2000)
For Research Use Only. Not for use in diagnostic procedures.