T1 Cartridges
Specifications for T1 Cartridges
Application: The cartridges contain a unique solid phase extraction (SPE) resin that binds a wide range of fluorescently labeled glycans and allows purification of these from labeling reagents. They are compatible with many leading vacuum manifold SPE handling systems. Labelled glycans can be N or O-link type.
Description: For post-labeling purification of fluorophore and chromophore labeled glycans. Suitable for cleanup after glycan labeling with 2-AB (2-aminobenzamide) and 2-AA (2-aminobenzoic acid).
Binding capacity: Each cartridge can typically bind glycans released from up to 50-100 µg glycoprotein.
Number of Samples: T1 cartridges are designed for single use only.
Suitable Samples: A wide range of glycans can be purified. These include N-linked and O-linked type oligosaccharides.
BindingSelectivity: Essentially stoichiometric binding and elution for most complex glycan mixtures.
Storage: Store at room temperature in the dark. Protect from sources of heat, light, and moisture. The cartridges are stable for at least two years as supplied.
Shipping: The product can be shipped at ambient temperature.
Handling: Ensure that any glass, plasticware or solvents used are free of glycosidases and environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrate.
Safety: Please read the Material Safety Data Sheets (MSDS's) for all chemicals used.
All processes involving hazardous reagents should be performed using appropriate personal safety protection - eyeglasses, chemically resistant gloves (e.g. nitrile), etc. - and where appropriate in a laboratory fume cupboard.
For research use only. Not for human or drug use
Additional Reagents and Equipment Required
Reagents
Pure water (HPLC grade)
Acetonitrile (HPLC grade)
Binding solution – 96 % acetonitrile in water (v/v)
Wash solution – 96 % acetonitrile in water (v/v)
Elution solution – pure water
Equipment
Pipettes
0.5 μm or 0.2 μm microcentrifuge filters
Microcentrifuge
Introduction
T1 cartridges have been designed for purification of glycans post-fluorophore labeling. The cartridges are designed for use with vacuum manifolds or liquid handling robots. They can also be used as stand-alone devices where the user relies on gravity for washing and elution steps.
Timeline for Cleanup
T1 glycan cleanup procedure typically takes around 55 minutes using a vacuum manifold.
Procedure | Time |
1. Assemble the vacuum manifold | 02 min |
2. Preparation of T1 cartridges | 10 min |
3. Preparation of samples for cleanup | 05 min |
4. Apply the sample to T1 cartridges | 08 min |
5. Washing cartridges | 15 min |
6. Elution of labelled glycans | 15 min |
Total Time | 55min |
1）Wash and prime the cartridge
Prepare each T1 cartridge by washing with the following:
Reagent Volume (ml）
1st wash:Water 1
2nd wash: 96 % acetonitrile* 1
*Prime with 100% acetonitrile for O-glycan clean-up.
2）Prepare the glycan samples
Pipette 200 µL of 96% acetonitrile (100% acetonitrile for O-link glycans) into your fluorophore labeled sample (typically fluorophore labelling mix + glycan is a 5-10 µL volume of sample). Gently mix the sample by pipette action.
3）Apply the samples to the cartridge
Load each sample onto a primed cartridge. Without applying a vacuum allow the sample to settle into the cartridge for 5 minutes. Most of the solution should flow into the cartridge over this time. Next, apply a slow vacuum (approximately taking one minute) to drain the T1.
4）Wash off non-glycan contaminants
Wash with 1mL of 96% acetonitrile* by applying the solution to the top of the cartridge and allow a slow vacuum to pull the solution through over a period of a few minutes. Repeat with 2 additional washes of 1ml 96% acetonitrile*. *100% acetonitrile for O-link glycan clean-up.
5）Elute the glycans
Remove the waste reservoir and replace with a collection reservoir. Without applying a vacuum, elute fluorophore labeled glycans with 0.5 mL purified water (for extra caution 1 mL can be used to elute glycans where very large glycans are expected) by adding the water to the top of each cartridge and allowing about 15 minutes for the solution to pass through the cartridge and drain into the collection plate. After 15 minutes a gentle vacuum can be applied to the cartridges to ensure all eluant has been drained from the cartridge. Slow sample binding and elution from SPE cartridges generally ensures better sample to sample reproducibility.