The Ludger Exoglycosidase Clean-up Cartridges have been developed for the removal of exoglycosidase enzymes and other protein material following glycan digestion or sequencing. This will prevent contamination of HPLC columns during subsequent chromatographic analysis. The spin columns can also be used to remove exoglycosidases or other proteins before mass spectrometry analysis of glycans.

The exoglycosidase clean-up spin columns contain a specialized modified polyethersulfone membrane with a molecular weight cut-off of approximately 30 kDa. This product is compatible with any centrifuge equipped with a rotor suitable for Eppendorf tubes. Glycans pass through the membrane whilst proteins are retained on the membrane allowing separation of these two components.

A 96-well exoglycosidase clean-up plate is also available.

Number of Samples Each pack contains 6 spin columns

Amount of Sample Up to 500 μL per column

Centrifugal Force Up to 14,000 x g

Suitable Samples Unlabelled and fluorophore labelled (e.g. 2-AB, 2-AA or procainamide labelled) glycans released from glycoproteins or other sources and treated with exoglycosidase enzymes.

Exoglycosidase Clean-up Method
Apply the samples onto the spin column
Pipette the glycan sample onto the spin column membrane. Wash out each sample vial with 100 μL of water and add this to the spin column membrane. Close the vial and centrifuge for 3 minutes at 10,000 x g or until the liquid has all gone through the membrane.
The maximum centrifugal force used should not exceed 14,000 x g.

Wash the membrane of the spin column
Pipette 100 μL of water directly onto the spin column membrane to wash through any remaining sample. Close the vial and centrifuge for 3 minutes at 10,000 x g or until the liquid has all gone through the membrane.

Dry the glycans if necessary
At this stage glycan samples may be at sufficient concentration for their intended use. Alternatively you additionally concentrate the glycans in a vacuum centrifuge. We do not recommend applying heat at this stage. Long drying times under elevated temperature may lead to glycan desialylation.

Storage Store at room temperature. Protect from sources of heat, light, and moisture. When stored correctly, the products should be stable for 36 months from date of purchase.

Shipping The product should be shipped at ambient temperature.