One kit contains reagents to label up to 96 separate analytical samples per kit, in two sets of reagents (48 samples per set). Typically, 50 micrograms of glycoprotein is analyzed per sample.
Released monosaccharides have a free reducing terminus allowing fluorescent tagging with 2-AA (2-aminobenzoic acid).
The kit also contains the LudgerTag MonoMix, a monosaccharide standard containing glucosamine, galactosamine, galactose, mannose and fucose. Xylose is included for use as an internal standard.
There are two hydrolysis acids provided. 2 molar trifluoroacetic acid (2M TFA) and 6 molar hydrochloric acid (6M HCl). 2M TFA is good for releasing neutral monosaccharides but is less effective with the N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc) monosaccharides for which we recommend using 6M HCl. We recommend using these acids on separate replicates of your samples.
LudgerTag Release Reagents
Trifluoroacetic acid – 2M TFA
Hydrochloric acid – 6M HCl
LudgerTag Monosaccharide Standards
MonoMix monosaccharide standards – 10 nmols each of glucosamine, galactosamine, galactose, mannose and fucose
Xylose standard – 100 nmols – used as an internal reference standard for accurate quantification
LudgerTag Labeling Reagents
2-aminobenzoic acid (2-AA dye)
Dimethyl sulfoxide (DMSO)
sodium acetate/boric acid/methanol
HPLC -Quantitative Analysis of Labelled Monosaccharides
Column: We offer two choices of column for analysis of the labelled monosaccharides dependent on whether you are using HPLC or UHPLC systems in your laboratory. If using an HPLC, the LudgerSep R2 column (Cat No. LS-R2-4.6×150) gives very good separation of the seven main monosaccharides found in most N-link and O- link glycans. If you have an UHPLC system we recommend using the LudgerSep uR2 column (Cat No. LS- UR2-2.1×50) which can perform an 8 minute separation per sample. See figure to the side.
Solvents: The glycan analysis gradients in this guide are based on the following solvents:
Solvent A : purified water based solvent of 0.2 % butylamine (2 mL per litre), 0.5 % phosphoric acid 5 mL per litre), 1 % tetrahydrofuran (10 mL per litre) (henceforth called BPT solvent).
Solvent B : 50 % acetonitrile : 50 % solvent A
We have also tested acetonitrile free solvent systems. Please enquire for further details.