Following on from research performed by Ludger Ltd. (www.ludger.com) in collaboration with Leiden University Medical Centre, The Netherlands, Ludger has developed labelling kits containing a different and non-toxic reducing agent, 2-picoline borane (2-PB). This technology has now been patented and Ludger is delighted to offer two new kits for labelling glycans.
Previously, 2-AA and 2-AB labelling kits used sodium cyanoborohydride as a reducing agent during glycan labelling. This reagent is toxic so a fume hood should be used during handling.
The kits have been validated following ICH Q2(R1) guidelines and beta-tested in the field. Precision values were excellent, with CVs of <5% for peaks with relative areas over 5% and CVs of <8% for peak with relative areas less than 5%.(see Figure 1). The kits offer equivalent labelling efficiency as standard LudgerTag™ 2AA and 2AB labelling kits containing sodium cyanoborohydride (Figure 2).
The kit contains two sets of the following reagents (12 samples per kit) supplied in glass ampoules sealed under pure nitrogen:
- 22-aminobenzoic acid (2-AA dye)
- Dimethyl sulfoxide (DMSO)
- Acetic acid
- Picoline borane
One kit contains reagents to label up to 24 separate analytical samples per kit.
The LudgerTag™ labeling procedure takes 2 hours with just 1 hour for the actual labelling incubation.
|Transfer samples to reaction tube and dry||30 min|
|Add water to samples||15 min|
|Make up and add labeling reagent||15 min|
|Incubate samples with reagent||1 hour|
|Total Time||2 hours|
- Anumula KR, Dhume ST. (1998)
High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid.
Glycobiology 8: 685-694
- Bigge JC, Patel TP, Bruce JA, Goulding PN, Charles SM, Parekh RB. (1995)
Nonselective and efficient fluorescent labeling of glycans using 2- amino benzamide and anthranilic acid.
Anal Biochem 230: 229-238