Permethylation kit (without methyl iodide)
Permethylation kit (without methyl iodide)
- Product Code: LT-PERMET-VP96
- Size 96 samples
- Ex Tax: £576
LudgerTag Permethylation of Glycans
Analysis of glycans by MALDI-MS provides mass composition data and is faster than UHPLC techniques (on average one sample can take as little as 1 minute for data acquisition), but sialylated glycans are usually more difficult to analyse because the sialic acid moieties are labile. Permethylation is a popular technique for the derivatisation of carbohydrates for MALDI-MS detection, as it enhances ionization efficiency, stabilizes the sialic acids and aids linkage analysis studies. However, conventional techniques used for permethylation are slow, labour intensive and therefore not suited to high throughput sample analysis.
We have developed a simple to use LudgerTag kit for permethylating glycans, LT-PERMET-VP96. The kit can be used to process 1-96 samples using either a manual method or an automated method that has been adapted to a liquid handling robot. It is suitable for N- or O-glycans and once permethylated, they can be analysed by MALDI-TOF-MS or LC-MS.
We have validated this technology and demonstrated that glycoprofiling data is reliable and comparable to the gold standard HILIC UHPLC method. This technology has been used for a range of high throughput studies at Ludger (and using the automated permethylation sample preparation time is under 5.5 hours for 96 samples).
Principles - Quick Links:
- Introduction: Why Permethylate?
- Ludger Glycan Permethylation Kit
- Manual/Automated Workflows
- Examples of how we're using LT-PERMET-96 at Ludger
- How to start using the Ludger Permethylation Kit
Posters & Publications
Shubhakar A, Kozak RP, Reiding KR, Royle L, Spencer DI, Fernandes DL, Wuhrer M.
Analytical Chemistry. 2016 Sep 6. 88(17):8562-9. doi: 10.1021/acs.analchem.6b01639. Epub 2016 Aug 15.
Shubhakar A, Spencer D, Wuhrer M, Fernandes DL
Presented at the 38th Annual Congress of Pharmacology Society of Chile
Castro, Chile. Nov 26-29th 2016
Reliability of a semi-automated, high throughput (HT) MS based glycomics system for discovery of glycan biomarkers and Quality by Design (QbD) studies of glycoprotein therapeutics
Shubhakar A, Spencer DI, Wuhrer M, Fernandes DL
Presented at Gordon Research Conference
Lucca, Italy. March 2015
Shubhakar A, Kozak RP, Badia-Tortosa C, Gardner RA, Royle L, Wuhrer M, Spencer DI, Fernandes DL
Presented at PEGS 2014: Glycosylation Analysis for Biopharmaceuticals Workshop
Boston, United States. May 2014
Optimisation of drug glycosylation supported by an automated high throughput permethylation MALDI-MS method
Shubhakar A, Spencer DI, Kozak RP, Velez-Suberbie L, Bracewell DG, Fernandes DL, Wuhrer M
Presented at ICS: 27th International Carbohydrate Symposium
Bangalore, India. January 2014
For the permethylation of free glycans. Kit sufficient for 96 samples. Without Methyl Iodide (MeI)
Due to shipping restrictions of dangerous goods this kit does not include methyl iodide (synonym: iodomethane). When ordering LT-PERMET-VP96 we recommend purchasing methyl iodide with purity ≥99.0% (GC) from your local chemical provider. CAS #:74-88-4
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
View product documentation
Application: This kit is used for the permethylation of glycans. Permethylation stabilizes glycan sialic acids for MALDI mass spectrometry analysis, and aids linkage analysis studies. The kit can be used with glycans released from glycoproteins.
Description: This kit contains reagents for the permethylation of glycans.Note: This kit does not include methyl iodide / Iodomethane (see shipping section for details)
Number of Samples: Sufficient for up to 96 samples.
Amount of Sample: Up to 1 μg of released glycans.
Suitable Samples: Any unlabelled purified glycans, released from glycoprotein by either PNGaseF, PNGase A, beta-elimination or hydrazinolysis.
Storage Store at 2-8°C.
Shipping: The product should be shipped between 2-8°C. 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
Safety: For research use only. Not for human or drug use. Please read the Safety Data Sheet (SDS’s) for all chemicals used. All processes involved permethylation reagents should be performed using appropriate personal safety protection – eyeglasses, chemically resistant gloves and where appropriate in a laboratory fume cupboard.
The kit contains the following items:
|LT-PERMET-PLATE-96||96 well Permethylation Plate||1 Plate|
|LT-PERMET-DMSO-96||dimethyl sulfoxide (DMSO)||40 mL|
|LT-PERMET-DCM-96||dichloromethane (DCM)||60 mL|
|LT-PERMET-LID-96||96 well Plate Lid||1 Lid / Mat|
|LT-BALANCE-PLATE||96 well Balance Plate||1 Plate|