Chunk #50 — MATERIALS AND METHODS — Immunoblotting of integrin β1 and Flag-Sdc4

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Musculocontractural Ehlers-Danlos syndrome and neurocristopathies: dermatan sulfate is required for Xenopus neural crest cells to migrate and adhere to fibronectin.

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Cleared lysates from the epidermis-enriched explants of 12 uninjected embryos were lysed in MBS, 1% Triton X-100, 5 mM EDTA and proteinase inhibitors for 30 min on ice, centrifuged at 10,000 g for 30 min at 4°C and dialyzed with 10 mM sodium acetate pH 7.0. The protein content was measured using the Bradford method (Bio-Rad), with BSA as a standard. Equal protein amounts were treated with lyases as described above.