Alpha-GalCer is an immunostimulant. It is a potent stimulator of natural killer T (NKT) cells and a specific ligand of the lipid-binding MHC class I-like protein CD1d in human and mouse NKT cells.
It protects against LPS-induced shock and is also a potent antitumor compound.
Synthetic. Originally isolated from the marine sponge Angelas mauritianus.
Solubility & Handling
Soluble in Tween20 or PBS for cell culture use. Also Soluble in pyridine (not suitable for cell culture). Aliquot into glass vials.
This compound is is inherently an extremely hydrophobic molecule and is insoluble in water, methanol, ethanol and other organic solvents. It is very slightly soluble in tetrahydrofuran and pyridine. Almost all methods for solubilizing this material in primarily aqueous media will contain at least some detergent and somewhat cloudy solutions may be produced which are still acceptable to use.
Some example protocols are as follows:
Tween20 & Sodium Chloride, Heat/Sonicate
Dissolve α-GalCer in 0.5% Tween20, 0.9% NaCl solution
Heat to 85°C until solution turns cloudy
Take out of water bath und keep at RT
Solution will become clear quickly
If particles are left, sonicate (important: sonication has to be performed in glass vial, not in a plastic Eppendorf tube) and filter afterwards with 2 µm filter
Tween20 & PBS, Heat/Sonicate
Dissolve the compound (0.2mg/ml) in PBS containing 0.5% Tween20 (warm and sonicate for 2 hrs. at 37°C)
The solution can be more like a suspension than a clear solution
If particles are left, warm up and sonicate (important: sonication has to be performed in glass vial, not in a plastic Eppendorf tube)
For sonication no interval is recommended, just use a conventional water bath sonicator at standard amplitude
Heating and sonication should be done immediately prior to every use.
Dissolve the compound in anyhdrous DMSO at a concentration of 1mg/ml with heating at 80°C for several minutes
Sonication for 2 h
The solution of 1mg/ml in DMSO can be further diluted in PBS
Please note the compound will likely precipitate out when added into aqeous media. It is therefore recommended that the aqueous media the DMSO stock solution will be diluted in should contain 10% serum/ BSA and the DMSO stock solution should be diluted no more than 1/100
Further heating/ vortexing/sonication may be required to dissolve any remaining precipitate which may take time and need to be repeated