Isolation and analysis of sugar nucleotides using solid phase extraction and fluorophore assisted carbohydrate electrophoresis
| Autor: | Liping Tian, Suzy A. A. Comhair, James Hiznay, Jarrod W. Barnes, Jacqueline Loftis, Raed A. Dweik, Mark E. Lauer |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Electrophoresis Carbohydrate Clinical Biochemistry GlcUA glucuronic acid UDP uridine diphosphate Nucleotide sugar High-performance liquid chromatography Man Mannose 03 medical and health sciences chemistry.chemical_compound Capillary electrophoresis GalNAc N-acetylgalactosamine TEMED N′ N′ N′N′-tetramethylenediamine Biochemistry Genetics and Molecular Biology Nucleotide AMAC 2-aminoacridone Sugar nucleotides Solid phase extraction Sugar ComputingMethodologies_COMPUTERGRAPHICS GDP guanosine diphosphate NeuAc sialic acid chemistry.chemical_classification Chromatography Chemistry CMP cytosine monophosphate TEAA triethylamine acetate APS ammonium persulfate GlcNAc N-acetylglucosamine Sugar nucleotide analysis by SPE and FACE FACE fluorophore assisted carbohydrate electrophoresis Gal galactose Medical Laboratory Technology 030104 developmental biology Biochemistry SPE solid phase extraction HPLC high performance liquid chromatography Face HPLC Fluorophore-assisted carbohydrate electrophoresis |
| Zdroj: | MethodsX |
| ISSN: | 2215-0161 |
| DOI: | 10.1016/j.mex.2016.03.010 |
| Popis: | Graphical abstract The building blocks of simple and complex oligosaccharides, termed sugar nucleotides, are often overlooked for their role in metabolic diseases and may hold the key to the underlying disease pathogenesis. Multiple reasons may account for the lack of analysis and quantitation of these sugar nucleotides, including the difficulty in isolation and purification as well as the required expensive instrumentation such as a high performance liquid chromatography (HPLC), mass spectrometer, or capillary electrophoresis. We have established a simple yet effective way to purify and quantitate sugar nucleotides using solid phase extraction (SPE) chromatography combined with fluorophore assisted carbohydrate electrophoresis (FACE). The simplicity of use, combined with the ability to run multiple samples at one time, give this technique a distinct advantage over the established methods for isolation and analysis of sugar nucleotides from cell culture models. • Sugar nucleotides can be easily purified with solid phase extraction chromatography. • FACE can be used to analyze multiple nucleotide sugar extracts with a single run. • The proposed method is simple, affordable, and uses common everyday research labware. |
| Databáze: | OpenAIRE |
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