Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1
Autor: | Rita Martins, Norman J. Dovichi, Colin D. Whitmore, Monica M. Palcic, Guido Tettamanti, Ole Hindsgaul, E. Andreas Larsson, Ulf Olsson, Ronald L. Schnaar |
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Rok vydání: | 2007 |
Předmět: |
endocrine system
Sialyltransferase Neuraminidase G(M2) Ganglioside G(M1) Ganglioside Biochemistry Article Analytical Chemistry Acylation Capillary electrophoresis Gangliosides Animals G(M3) Ganglioside Fluorescent Dyes chemistry.chemical_classification Ganglioside Chromatography biology Rhodamines Organic Chemistry Electrophoresis Capillary General Medicine Reference Standards Fluorescence carbohydrates (lipids) Enzyme chemistry biology.protein Cattle lipids (amino acids peptides and proteins) Naked eye |
Zdroj: | Carbohydrate Research. 342:482-489 |
ISSN: | 0008-6215 |
Popis: | Ganglioside GM1 and its seven potential catabolic products: asialo-GM1, GM2, asialo-GM2, GM3, Lac-Cer, Glc-Cer and Cer, were labeled with tetramethylrhodamine (TMR) to permit ultra-sensitive analysis using laser-induced fluorescence (LIF) detection. The preparation involved acylation of the homogenous C(18)lyso-forms of GM1, Lac-Cer, Glc-Cer and Cer with the N-hydroxysuccinimide ester of a beta-alanine-tethered 6-TMR derivative, followed by conversion of these labeled products using galactosidase, sialidase, and sialyltransferase enzymes. The TMR-glycolipid analogs produced are detectable on TLC down to the 1 ng level by the naked eye. All eight compounds could be separated within 4 min in capillary electrophoresis where they could be detected at the zeptomole (ca. 1000 molecule) level using LIF. |
Databáze: | OpenAIRE |
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