Analysis of Aggrecan Glycanation by Western Blot in Cell Culture.

Autor:	Gramegna Tota C; Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, Pavia, Italy., Leone A; Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, Pavia, Italy.; University School for Advanced Studies Pavia, IUSS Pavia, Pavia, Italy., Rossi A; Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, Pavia, Italy. antrossi@unipv.it., Paganini C; Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, Pavia, Italy.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2619, pp. 141-151.
DOI:	10.1007/978-1-0716-2946-8_11
Abstrakt:	Several experimental protocols are available to study the synthesis and secretion of proteoglycans in health and diseases, but there are few methods to analyse the intracellular processing of these macromolecules. We report a western blot analysis on medium and cell layer of primary chondrocyte culture to determine the glycanation status of aggrecan. Using a specific antibody against the aggrecan core protein and digesting an aliquot of sample with chondroitinase ABC, it is possible to analyse the whole aggrecan macromolecule and the core protein in order to evaluate defects in aggrecan glycanation. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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