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. 2018 Jul 17;28(10):786–801. doi: 10.1093/glycob/cwy057

Fig. 9.

Fig. 9.

ADAMTS-4 (aggrecanase-1) treatment of human airway Siglec-8 ligands shifts their electrophoretic migration. Siglec-8 ligands were extracted from human trachea, purified by sequential size exclusion followed by Siglec-8 affinity chromatography and subjected to ChABC, aggrecanase or both enzymes. Samples were resolved on 1.5% acrylamide, 2% agarose composite gels, blotted to PVDF membranes and probed with precomplexed Siglec-8-Fc or anti-aggrecan antibody (7D4) as indicated. Lanes (all enzyme treatments 16 h at 37°C): (1) incubation without enzyme; (2) aggrecanase (0.3 mU/mL); (3) aggrecanase (0.3 mU/mL) plus ChABC (0.5 U/mL); (4) ChABC (0.5 U/mL). S8-1M and S8-600K were resolved on 1.5% acrylamide, 2% agarose composite gels with migration positions of HiMark molecular weight markers shown; S8-250K was resolved on 3% acrylamide, 2% agarose composite gels with migration positions of SeeBlue Plus2 molecular weight markers shown.