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  • Separation and Purification of Glycosaminoglycans (GAGs) from Caenorhabditis elegans.

Separation and Purification of Glycosaminoglycans (GAGs) from Caenorhabditis elegans.

Bio-protocol (2017-08-05)
Tabea Dierker, Lena Kjellén
摘要

The nematode Caenorhabditis elegans is a popular model organism for studies of developmental biology, neurology, ageing and other fields of basic research. Because many developmental processes are regulated by glycosaminoglyans (GAGs) on cell surfaces and in the extracellular matrix, methods to isolate and analyze C. elegans GAGs are needed. Such methods have previously been optimized for other species such as mice and zebrafish. After modifying existing purification protocols, we could recently show that the nematodes also produce chondroitin sulfate, in addition to heparan sulfate, thus challenging the view that only non-sulfated chondroitin was synthesized by C. elegans. We here present our protocol adapted for C. elegans. Since the purification strategy involves separation of non-sulfated and sulfated GAGs, it may also be useful for other applications where this approach could be advantageous.

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Sigma-Aldrich
琼脂糖, BioReagent, Molecular Biology, low EEO
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
胆固醇, Sigma Grade, ≥99%
Sigma-Aldrich
软骨素酶 ABC 来源于普通变形杆菌, BSA free, lyophilized powder, specific activity 50-250 units/mg protein