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

Productα-GalCer unloaded
MBL code no.
α-GalCer loaded
MBL code no.
Human CD1d Tetramer-PETS-HCD-1TS-HCG-1
Human CD1d Tetramer-APCTS-HCD-2TS-HCG-2
Mouse CD1d Tetramer-PETS-MCD-1TS-MCG-1
Mouse CD1d Tetramer-APCTS-MCD-2TS-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 µl 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.


  1. Stephane S, et al., J. Immunol. Methods 268: 107-121 (2002)
  2. Matsuda JL, et al., J. Exp. Med. 192: 741-754 (2000)
  3. Benlagha K, et al., J. Exp. Med. 191: 1895-1903 (2000)

For Research Use Only. Not for use in diagnostic procedures.

Learn more about the other types of MHC Molecules