Cleanup of N-labelled glycans
Cleanup of N-labelled glycans
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Micro-SPE Techniques (MiniSpin
columns, 96-Well Spin plates)
Cleanup of N-labelled glycans in NEB Rapid PNGase F buffer from 2AB, or procainamide reagent, NaCNBH4, DMSO, and acetic acid labelling solution.
With thanks to Paula Magnelli, et al., New England Biolabs
Note: These loading volumes are high but the cartridges are on a vacuum manifold so the excess liquid is pulled through the cartridges.
HILIC cleanup (vacuum manifold) of MicroSpin HILIC (SEM HIL):
Condition SEM-HIL HILIC columns with 350µl Acetonitrile.
Add 350µl, 50mM ammonium formate, pH 4.4. Repeat two times.
Equilibrate with 350µl of a solution 90% Acetonitrile/10% 50mM ammonium formate, pH 4.4. Repeat two times
Bring glycan labeling reaction to 87-85% acetonitrile, mix well. Total volume 100-200µl
Load sample slowly into an equilibrated SPE column.
Wash with 350µl of a solution 90% Acetonitrile/10% 50mM ammonium formate, pH 4.4. Repeat five times
Spin at 3000 rpm 1 min to dry.
Elute with 50µl of 50mM ammonium formate, pH4.4: Let it enter by gravity, incubate for 2min. Spin 3000 rpm 2 min.
Overview of All Chemistries
and Configurations
General Operating Instructions:
General HILIC: Hydrophilic Interaction and Detergent Removal Instructions
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Last Updated: 08/20/21
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