Analysis of Aggrecan Glycanation by Western Blot in Cell Culture.

Chiara Gramegna Tota, Alessandra Leone, Antonio Rossi, Chiara Paganini

Methods in molecular biology (Clifton, N.J.) 2023

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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.

Citation

Chiara Gramegna Tota, Alessandra Leone, Antonio Rossi, Chiara Paganini. Analysis of Aggrecan Glycanation by Western Blot in Cell Culture. Methods in molecular biology (Clifton, N.J.). 2023;2619:141-151