FAQ: What are the best techniques to analyze N- and O-glycans?
Glycans are neutral or charged molecules with no intrinsic UV absorbance or autofluorescent emission. Therefore carbohydrate analysis requires dedicated instrumentation. Glycans released from proteins can be analyzed directly by HPAEC-PAD (High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection); or they are derivatized with fluorescent or methyl groups for subsequent analysis by capillary electrophoresis and/or liquid chromatography/mass spectrometry (LC/MS). Finally, abundant glycan samples can be separated by TLC, however the resolution is poor for complex mixtures and large molecules.
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