The GlykoPrep H Cleanup Module contains GlykoPrep HC solid-phase extraction spin cartridges packed with ~5 mg of Hypercarb™ porous graphitic carbon (PGC). These are designed for purification of glycans from non-carbohydrate material, including salts and proteins. When samples are loaded on the column, salts pass through, while glycans and protein adsorb onto the matrix. The glycans may then be selectively eluted from the column.
Note: these cartridges are not suitable for removing detergent.
*Not recommended for cleanup of InstantPC-labeled N-glycans, as these must be stored in Gly-X/InstantPC Eluent with 10% ACN. Samples applied to HC cartridges must be aqueous or low organic (<5%).
Sample prep considerations:
• Glycan samples to be cleaned must be in an aqueous buffer or a buffer containing a low percentage (<5%) of organic solvent.
• If the sample contains organic solvent, dilute with water until the organic solvent content is less than 5% by volume. For example, dilute a 5 μl 2-AB labeling reaction with 95 μl water prior to loading.
• Samples should not contain detergents such as SDS.
• Suggested starting upper limit for the HC cartridges is ~50 μg glycans. The binding capacity for specific glycoproteins & glycans may need to be determined. Minimum glycan load will depend on the sensitivity of the analytical methods used.
The H Cleanup Module requires minimal equipment. All you need is a centrifuge with a deep-well plate rotor (minimum 44 mm height) and a Starter Labware Kit (AM200).
Size: 1 ea
Product Code: GS24-HC
Q: Do you have a protocol for cleanup of glycans released by hydrazinolysis
using the HC cartridges?
A: No, but Hypercarb material has been used to clean up glycans released using our Glycan Hydrazinolysis Kit .
Q: Is it possible to preferentially elute neutral N-glycans vs. sialylated N-glycans from the HC resin?
A: Developmental work at ProZyme indicates that neutral glycans can be eluted alone using an eluent of 25% acetonitrile. Then, sialylated glycans may be eluted with 25% acetonitrile with 0.1% TFA. Using this two-step elution, neutral and sialylated glycans may be fractionated. Please contact ProZyme for further details.
Q. Do you have a larger version of the HC cartridge?
A. The GlycoClean H Cartridge from ProZyme (product code GKI-4025) is a larger gravity flow cartridge packed with 200 mg of Hypercarb.
1. Turyan I, Hronowski S, Lyubarskaya Y. Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. Anal Biochem. 2014 Feb 1;446:2836