ALEXIS Biochemicals: Isoglobotrihexosylceramide

ALX-306-028

Revised 05-May-08

Isoglobotrihexosylceramide
SYNONYMS	iGb3
PRODUCT LINE	Immunology
PRODUCT CATEGORY	Natural Killer T [NKT] Cells / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-306-028-C100		100 µg	85.00 USD
ALX-306-028-M001		1 mg	455.00 USD

Product Specification

FORMULA:	C₆₂H₁₁₇NO₁₈
MW:	1164.6
PURITY:	≥95% (¹H-NMR)
APPEARANCE:	White to off-white powder.
SOLUBILITY:	Soluble in pyridine or chloroform:methanol (2:1).
SHIPPING:	SHIPPED ON BLUE ICE
SHORT TERM STORAGE:	+4°C
LONG TERM STORAGE:	-20°C

Product Description

Lysosomal glycosphingolipid proposed to be the natural ligand of NKT cells. Stimulates natural killer T (NKT) cells similar to α-GalCer. For negative control compound see globotrihexosylceramide (Gb3) (Prod. No. ALX-306-030).

Product Specific Literature References

[1] Lysosomal glycosphingolipid recognition by NKT cells: D. Zhou, et al.; Science 306, 1786 (2004) Abstract
[2] Synthesis and biological evaluation of alpha-galactosylceramide (KRN7000) and isoglobotrihexosylceramide (iGb3): C. Xia, et al.; Bioorg. Med. Chem. Lett. 16, 2195 (2006) Abstract
[3] Chemoenzymatic Syntheses of iGb3 and Gb3: Q. Yao et al.; Org. Lett. 8, 911 (2006) Abstract

General Information

BACKGROUND/TECHNICAL INFORMATION

Solubilization:
Our recommendation is to dissolve product in chloroform: abs. methanol 2:1. Aliquot into glass tubes, dry and store at 4°C. Resuspended in abs. methanol (at concentrations not higher than 1%) prior to cell assays. Note: The use of DMSO may abolish the iGb3 activity and should be avoided [2].

For expanding human NKT cells from freshly isolated peripheral blood mononuclear cells (PBMC):
Mix iGb3 at 100ng/ml with PBMC and culture for 4-5 days.

For the stimulation of mouse NKT cells:
The bone marrow derived dendritic cells (BMDC) is the most efficient cell type that present iGb3. Use 50’000-250’000 of BMDCs pulsed overnight with 100ng/ml of iGb3 and 50’000 freshly purified NKT cells as a responder. Use tetramer sorting to purifiy the NKT cells from spleen and before mixing with BMDCs, rest them in culture medium for 12 hours and then wash with medium to avoid a high background. MACS sorting technique is not recommended. Alternatively, NKT cell hybridomas can be used to detect the ligand. (For reference see: http://home.uchicago.edu/~dzhou/Testing%20iGb3%20ligand.htm)