columns and vacuum dried. The exhaustive depolymerization was accomplished by dissolving one sixth of each sample in 10 μl water, followed by adding 8 μl 100 mM Tris/HCl pH 7.45 2 μl 1 M ammonium acetate, 20 mU of chondroitinase ABC, 10 mU chondroitinase AC-I, and 1.25 mU chondroitinase B. In a distinct incubation, 2 mU of chondroitinase B only was added to the GAG samples, in the presence of 0.1% BSA and 1 μM CaCl2 to specifically cleave at IdoA of DSs. In both cases, the digest solutions were incubated at 37 °C, and after two hours, another aliquot of lyases was added.
