Rapid detection of extended ganglio-series gangliosides with terminal GalNAc beta 1-4Gal sequence on high performance thin layer chromatography plates.
The mouse monoclonal antibody 2D4, which recognizes the terminal GalNAc beta 1-4Gal-disaccharide of GgOse3Cer and GgOse5Cer, was used for the detection of ganglio-series gangliosides. The method involves separation of gangliosides on thin layer chromatography plates, followed by silica gel fixation, Arthrobacter ureafaciens neuraminidase treatment and final immunostaining of desialylated gangliosides with the monoclonal antibody 2D4. Both neuraminidase and the hybridoma 2D4 producing the specific monoclonal antibody are commercially available and therefore accessible to all researchers working in this field. Gangliosides from mouse T lymphocytes and the mouse T cell lymphoma YAC-1 have been used as examples. This technique may be used for fast screening of gangliosides with the GgOse5Cer core structure which have been described as T cell markers, antigens in human neuronal disease and receptors for certain pathogenic bacteria.
Müthing
J
J
Institut für Biotechnik Tierischer Zellen, Universität Bielefeld, FRG.
Ziehr
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D001923:Brain Chemistry; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D005732:Gangliosides / Q000032:analysis*; D008223:Lymphoma / Q000032:analysis; D051379:Mice; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009439:Neuraminidase; D013601:T-Lymphocytes / Q000032:analysis
D000911:Antibodies, Monoclonal; D005732:Gangliosides; C064399:gangliosidepentaosylceramide; D009439:Neuraminidase