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Glycosylation

Glycoprofile™ Labeling Kits

The GlycoProfile™ Labeling Kits are designed for efficient labeling of N-linked, O-linked and glycosylphosphatidylinositol (GPI) anchored glycans using your choice of 2-aminobenzamide (2-AB) or 2-aminobenzoic acid (anthranilic acid; 2-AA) 1,2. These small fluorophores increase the spectral absorption of glycans, improving detection in HPAE (high-performance anion exchange chromatography),3,4,5,6 HPLC4,5,6,7, and ESI-MS (electrospray mass spectroscopy) methods. Binding is robust and the resulting dye-glycan conjugates are stable, with no degradation during post-labeling analysis. 2-AB is recommended for downstream analysis by HPAE, HPLC and ESI-MS, although it is slightly less sensitive than 2-AA. 2-AA is recommended for SDS-PAGE, and is also suitable for HPAE, HPLC and ESI-MS.


Labeling of carbohydrates with 2-AB


Labeling of carbohydrates with 2-AB Image

Features and Benefits

  • Each kit is complete, containing 2-AA or 2-AB dye, reductant and solvents.
  • Sufficient for the preparation of up to 36 samples of mixed glycans containing from 100 picomoles to 50 nanomoles of purified glycans. With a single pure glycan, as little as 5 picomoles may be labeled and detected in subsequent HPLC analysis.

 

Note: Labeling of glycans with 2-AB is covered under US Patent No 5,747,347 and its foreign equivalents.


GlycoProfile™ Glycan Clean-Up Cartridges are recommended for use with the GlycoProfile Labeling kits for post-labeling cleanup. Glycans adsorb to the membrane while excess dye, mono- and disaccharides and other hydrophobic contaminants pass through. Purified labeled glycans are eluted with water and are ready for subsequent enzymatic digestion or analysis.


Ordering Information

Product Code Product Name Unit Technical Bulletin
PP0530 GlycoProfile 2-AA Labeling Kit 1 kit
PP0520 GlycoProfile 2-AB Labeling Kit 1 kit
G4548 GlycoProfile Glycan Clean-up Cartridges 3 each
6 each
12 each

N/A


References:

  1. Bigge, J.C., et al., Non-selective and efficient fluorescent labeling of glycans using 2-aminobenzamide and anthranilic acid, Anal. Biochem., 230, 229-238 (1995).
  2. Hardy, M.R., Glycan labeling with the fluorophores 2-aminobenzamide and anthranilic acid, Techniques in Glycobiology, Dekker, Inc. NY (1997).
  3. Delaney, J., and Vouros, P., Liquid chromatography ion trap mass spectrometric analysis of oligosaccharides using permethylated derivatives, Rapid Commun. Mass Spectrom., 15(5), 325-334 (2001).
  4. Anumula, K.R., and Dhume, S.T., High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivitization with highly fluorescent anthranilic acid, Glycobiology, 8, 685-694 (1998).
  5. Townsend, R., et al., Multimode high-performance liquid chromatography of fluorescently labeled oligosaccharides from glycoproteins, Anal. Biochem., 239, 200-207 (1996).
  6. Royle, L., et al., An Analytical and Structural Database Provides a Strategy for Sequencing O-Glycans from Microgram Quantities of Glycoproteins, Anal. Biochem., 304, 70-90 (2002).
  7. Guile, G.R., et al., A rapid and high-resolution high-performance liquid chromatographic method for separating glycans mixtures and analyzing oligosaccharide profiles, Anal. Biochem., 240, 210-226 (1996).
  8. Rudd, P.M., et al., Glycoproteins: rapid sequencing technology for N-linked and GPI Anchor glycans, Biotechnol. Genet. Eng. Rev., 16, 1-21(1999).