LudgerClean 96孔外切糖苷酶酶解后纯化板 The Ludger Exoglycosidase Clean-up Plate have been developed for the removal of enzymes and other protein material following glycan digestion or sequencing. This will prevent contamination of HPLC columns during subsequent chromatographic analysis. The 96-well plate can also be used to remove exoglycosidases or other proteins before mass spectrometry analysis of glycans.

The exoglycosidase clean-up plate contain a specialized modified polyethersulfone membrane with a molecular weight cut-off of approximately 30 kDa. This product is compatible for use in a vacuum manifold system (e.g. Ludger LV system) with any centrifuge equipped with a 96-well plate rotor. Glycans pass through the membrane whilst proteins are retained on the membrane allowing separation of these two components.

A exoglycosidase clean-up spin column is also available.

Number of Samples Up to 96 samples

Amount of Sample Up to 350 μL per column

Centrifugal Force Up to 1500 x g

Operating vacuum 10-20 in Hg (approx. 0.35-0.7 bar)

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 exoglycosidase clean-up plate
Place a 96-well collection plate (LP-COLLPLATE-2ML) inside the vacuum manifold. Assemble the manifold with the post-exoglycosidase clean-up plate on top ensuring that the collection plate is in- line with the wells (if using centrifugation instead, place the clean-up plate directly on top of the collection plate).
Ensure that the distance between the collection plate and the manifold top is as small as possible to reduce the gap between the clean-up plate and the collection plate and prevent sample cross- contamination.

Pipette the glycan samples into the post-exoglycosidase clean-up plate wells. Wash out each sample vial with 100 μL of water and add this to the clean-up plate wells. Apply a vacuum and adjust to between -0.3 and -0.5 bar until the liquid has all gone through the wells. Open the tap to release the vacuum.
Apply a vacuum and adjust to between -0.3 and -0.5 bar until the liquid has all gone through the wells. Open the tap to release the vacuum.

Wash the wells of the clean-up plate
Pipette 100 μL of water into each well to wash through any remaining sample. Apply a vacuum and adjust to between -0.3 and -0.5 bar until the liquid has all gone through the wells. Open the tap to release the vacuum.

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.