Abstract
A modified method for measuring the activities of glycosyltransferases using highperformance thin-layer chromatographic (HPTLC) plates is described. In this method a mixture of glycosphingolipid acceptors were chromatographed on a silica gel coated plate and were incubated with the enzyme solution and an appropriate radio-active sugar nucleotide donor. Following incubation, the plate was washed with a buffer and the radiolabeled reaction products on the plate were scrapped off and the radioactivities determined using a liquid scintillation counter. Alternatively, the plate could be exposed to an X-ray film to reveal the radioactive products. This method was used to assay the activities of rat brain cytidine 5’-monophosphate-N-acetylneuraminic acid: LacCer-, GM3-, GM1-, or GD3-sialyltransferases. We found that this method was sensitive, fast and reliable and was capable of assaying.
Original language | English (US) |
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Pages (from-to) | 2771-2781 |
Number of pages | 11 |
Journal | Journal of Liquid Chromatography |
Volume | 13 |
Issue number | 14 |
DOIs | |
State | Published - Aug 1 1990 |
Externally published | Yes |
ASJC Scopus subject areas
- Molecular Medicine