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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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
Cholesterol, Sigma Grade, ≥99%
Sigma-Aldrich
Agarose, BioReagent, Molecular Biology, low EEO
Sigma-Aldrich
Chondroitinase ABC from Proteus vulgaris, BSA free, lyophilized powder, specific activity 50-250 units/mg protein