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Permethylation Kit

This kit is used for the permethylation of glycans. Permethylation stabilizes sialic acids for MALDI mass spectrometry analysis, and aids linkage analysis studies (MALDI-MS, ESI-MS).

– Improves and enhances ionization efficiency of glycans on mass spec
– Increased glycan hydrophobicity enables LC-MS analysis
– Relative and absolute quantitation can be performed
– Stabilizes the sialic acid moieties
– Easier determination of branching and glycosidic linkage positions
– Enables detection of both neutral and acidic glycans in positive ion mode

Photo Guide of the Procedure
View product documentation as PDF
View Certificate of Analysis as PDF

Part Number: LT-PERMET-VP96

$989.00

Kit Contents



96 well Permethylation plate1 Plate
Dimethyl sulfoxide (DMSO)40.0 ml
Dichloromethane (DCM)60.0 ml
96 well Balance Plate1 Plate
96 well Plate Lid1 Lid / Mat

The plate is scalable and can be used for any number of samples between 1 and 96. Store the unused kit reagents and the unused plate wells in the recommended storage conditions to enable their use for further sample preparation at a later date.

Note: Due to shipping restrictions of methyl iodide/iodomethane, we are unable to provide this component with the kit. Therefore, we recommend purchasing iodomethane with purity ≥99.0% (GC) from your local chemical provider. Iodomethane Synonym: methyl iodide CAS Number: 74-88-4

 

Timeline for the Procedure

Step 1: Permethylation of samples
a. Preparation of glycansAs required
b. Addition of DMSO and sample to plate15 minutes
c. Incubation15 minutes
d. Addition of MeI to permethylation plate15 minutes
e. Incubation60 minutes

Step 2: Extraction of permethylated samples and storage
a. Extraction of samples3-4 hours
b. Drying permethylated samples10-30 minutes
c. Addition of 70% MeOH for storage10-20 minutes

 

References

A. A Ki-Irem; G. V. Avvakumov; I. V. Sidorova and A. Strel’Chyonok (1979) ‘Methylation analysis in glycoprotein chemistry’, Chromatography 180: 69-82

A. Dell; A. J. Reaseon; Kay-Hooi Khoo; M. Panico; A. McDowell and H. R. Morris (1994) ‘Mass Spectrometry of Carbohydrate-containing Biopolymers’, Methods in Enzymology 230: 108-132

I.Ciucanu; C. E. Costello (2003) ‘Epmination of Oxidative Degradation during the per-O-Methylation of Carbohydrates’, J. Am. Chem. Soc. 125: 16213-16219

I.Ciucanu; (2006) ‘Per-O-Methylation reaction of structural analysis of carbohydrates by mass spectrometry’ Analytica Chimica Acta 576: 147-155

I.Ciucanu; R. Caprita (2007) ‘Per-O-Methylation of neutral carbohydrates directly from aqueous samples for gas chromatography and mass spectrometry analysis’, Analytica Chimica Acta 585: 81-85

Shubhakar, A., Kozak, R.P., Reiding, K.R., Royle, L., Spencer, D.I.R., Fernandes, D.L., Wuhrer, M.: Automated High-Throughput Permethylation for Glycosylation Analysis of Biologics using MALDI- TOF-MS. Analytical Chemistry. acs.analchem.6b01639 (2016).

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