Glycosaminoglycan Lyase Digestion

1. Chondroitinase ABC (Proteus vulgaris), (Seikagaku America), dissolved in water at 1.0 U/mL and stored in 10-^L aliquots at -80°C for up to 3 mo (see Notes 4-6).

2. 50 mM ammonium acetate, pH 7.0, freshly prepared.

3. Microcentrifuge tubes (1.5 mL capacity).

Chondroitinase
Fig. 2. Schematic representation of chondroitinase ABC cleavage in the glycosaminoglycan chain and the products generated from the interior and the nonreducing terminus.
Reductive Amination Saccharide

Fig. 3. Schematic representation of saccharide fluorotagging by reductive amination with 2-aminoacridone (AMAC) and sodium cyanoborohydride. In this reaction a Schiff's base is formed between the aldol group of the reducing saccharide and the amino group of a fluorotag, and this bond is then reduced to the stable secondary amine linkage to complete the formation of the fluorotagged products (26,30).

Fig. 3. Schematic representation of saccharide fluorotagging by reductive amination with 2-aminoacridone (AMAC) and sodium cyanoborohydride. In this reaction a Schiff's base is formed between the aldol group of the reducing saccharide and the amino group of a fluorotag, and this bond is then reduced to the stable secondary amine linkage to complete the formation of the fluorotagged products (26,30).

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