TY - JOUR
T1 - Quantification of gangliotetraose gangliosides with cholera toxin
AU - Wu, Gusheng
AU - Ledeen, Robert
N1 - Funding Information:
This work was supported by PHS Grant 2-ROI-NS-04834. We gratefully acknowledge the skillful assistance of Agostino Cervone and the helpful advice of Sarah Ehrlich-Rogozinski in developing the TLC overlay procedure.
PY - 1988/9
Y1 - 1988/9
N2 - A procedure is described for assay of GM1 and other gangliotetraose-type gangliosides at the picomole level. The gangliosides are adsorbed onto polystyrene microwells and treated with neuraminidase and then with cholera toxin B subunits conjugated to horseradish peroxidase. Color is developed and quantified spectrophotometrically. Omission of neuraminidase gives a measure of GM1 alone. Linearity was obtained between 0.5 and 3 pmol. This procedure was applied to ganglioside mixtures isolated fron neuro-2A neuroblastoma and PC12 pheochromocytoma cells. For the latter, an additional step involving reaction with fucosidase increased the yield of GM1 due to the presence of fucosylated gangliosides. Application of the same reagents as a TLC overlay procedure to the gangliosides from neuro-2A cells revealed the presence of GD1a, GD1b, and GT1b in addition to GM1, thus confirming the presence of a family of gangliotetraose gangliosides.
AB - A procedure is described for assay of GM1 and other gangliotetraose-type gangliosides at the picomole level. The gangliosides are adsorbed onto polystyrene microwells and treated with neuraminidase and then with cholera toxin B subunits conjugated to horseradish peroxidase. Color is developed and quantified spectrophotometrically. Omission of neuraminidase gives a measure of GM1 alone. Linearity was obtained between 0.5 and 3 pmol. This procedure was applied to ganglioside mixtures isolated fron neuro-2A neuroblastoma and PC12 pheochromocytoma cells. For the latter, an additional step involving reaction with fucosidase increased the yield of GM1 due to the presence of fucosylated gangliosides. Application of the same reagents as a TLC overlay procedure to the gangliosides from neuro-2A cells revealed the presence of GD1a, GD1b, and GT1b in addition to GM1, thus confirming the presence of a family of gangliotetraose gangliosides.
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U2 - 10.1016/0003-2697(88)90201-1
DO - 10.1016/0003-2697(88)90201-1
M3 - Article
C2 - 3189815
AN - SCOPUS:0023792691
SN - 0003-2697
VL - 173
SP - 368
EP - 375
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -