Preparation of Alexa Fluor 350-conjugated nonradioactive or 3H-labeled GM1 ganglioside derivatives with different ceramides
| Autor: | Hanna Domek, Natalia Fedoryszak, Tadeusz Pacuszka, Mirosława Panasiewicz |
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| Rok vydání: | 2009 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Electrospray Biophysics HL-60 Cells G(M1) Ganglioside Acetates Ceramides Tritium Hydrazide Biochemistry Aldehyde High-performance liquid chromatography chemistry.chemical_compound Humans Tissue Distribution Bovine serum albumin Molecular Biology Chromatography High Pressure Liquid Alexa Fluor chemistry.chemical_classification Chromatography biology Cell Membrane Fatty acid Cell Biology Thin-layer chromatography chemistry Chromones biology.protein |
| Zdroj: | Analytical Biochemistry. 385:168-170 |
| ISSN: | 0003-2697 |
| Popis: | Alexa Fluor 350 hydrazide (AF) was coupled to the aldehyde group at C-6 of terminal galactose of oxidized GM1 gangliosides containing different fatty acid residues (GM1s). The AF–GM1 hydrazones obtained were reduced with NaBH4 or [3H]NaBH4 and purified by high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). Final yields of AF–GM1s exceeded 30%, purity was better than 97%, and radiochemical purity of 3H-labeled AF–GM1s was more than 94.5%. Structures of AF–GM1s were confirmed by electrospray ionization–mass spectrometry (ESI–MS). When added to HL-60 cell culture media, more than 81.6 or 78.9% of the AF–[3H]GM1s were taken up by cells in a bovine serum albumin- or trypsin-resistant manner, respectively. Approximately 70% of the AF–[3H]GM1s were recovered in HL-60 total plasma membrane fraction. |
| Databáze: | OpenAIRE |
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