The Procainamide Cleanup Plate is a 96 well membrane-bottom plate containing a specialized glycan binding membrane. This product is designed for use with the Velocity vacuum manifold or with other popular vacuum manifold systems. Excess procainamide label passes through the membrane whilst glycans are bound to the membrane allowing separation of these two components.

Number of Samples :Sufficient for up to 96 samples.

Volume of Sample: Up to 350 µL per well.

Suitable Samples: Procainamide labeled glycans. Typical samples would be procainamide labelled glycans from up to 100 µg of IgG glycoprotein.

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

Shipping: The product should be shipped at ambient temperature.

Procainamide Cleanup Method
Prepare the procainamide labeled glycan samples
Pipette 230 μL of acetonitrile into the procainamide labelled glycan samples (typically labelling mix + glycan + water is a 20 μL volume of sample). Gently mix the sample by pipette action.

Apply the samples to the Procainamide Cleanup Plate
Load each sample into a primed well of the plate. Without applying a vacuum allow the sample to settle into the cartridge for 5 minutes. Apply a vacuum and adjust to between -0.05 and -0.1 bar until the acetonitrile sample solution has all gone through the wells. Discard the waste.

Wash off non-glycan contaminants
Add 200 μL of acetonitrile to each well containing sample. Apply a vacuum and adjust to between -0.05 and -0.1 bar until the acetonitrile has all gone through the wells. Repeat with two additional washes of 200 μL of acetonitrile. Discard the waste.

Elute the labeled glycans
Place a 96 well collection plate (LP-COLLPLATE-2ML) inside the vacuum manifold. Assemble the manifold with the procainamide clean-up plate on top ensuring that the collection plate is in-line with the wells. Ensure that the distance between the collection plate and the manifold top is as small as possible to reduce the gap between the procainamide clean-up plate and the collection plate (spacers may be required to lift the collection plate).

Add 100 μL of water to each well containing sample. Apply a vacuum and adjust to between -0.05 and – 0.1 bar until the liquid has all gone through the wells. Open the tap to release the vacuum. Apply a higher vacuum (-0.5 bar) to remove as much remaining water from the wells (and underneath the membrane) as possible. Open the tap to release the vacuum. Repeat the higher vacuum (-0.5 bar) step for a second time.

Add another 100 μL of water into each well to wash through any remaining procainamide labelled sample. Apply a vacuum and adjust to between -0.05 and -0.1 bar until the liquid has all gone through the wells. Open the tap to release the vacuum. Apply a higher vacuum (-0.5 bar) to remove as much remaining water from the wells (and underneath the membrane) as possible. Open the tap to release the vacuum. Repeat the higher vacuum (-0.5 bar) step two additional times. These vacuum steps are done to remove as much remaining water from the wells (and underneath the membrane) as possible.

Dry down the samples
Samples can be stored for several hours at 4°C or for a longer period at -20°C. Samples can be stored in the collection plate, covered with a lid (LP-COLLPLATE-LID-96), or transferred to 0.5 mL centrifuge vials. Optionally, dry the samples down completely using a rotary speed vac (approximately 8 hours). We do not recommend applying heat at this stage. Only use a good quality vacuum centrifuge as long drying times or heat may lead to glycan desialylation.