How to detect and purify tenascin-C.
Fiche publication
Date publication
janvier 2018
Journal
Methods in cell biology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr OREND Gertraud
Tous les auteurs :
Giblin SP, Murdamoothoo D, Deligne C, Schwenzer A, Orend G, Midwood KS
Lien Pubmed
Résumé
The extracellular matrix molecule tenascin-C (TNC) was discovered over 30 years ago, and its tightly regulated pattern of expression since sparked keen interest in the scientific community. In adult tissues, TNC expression is restricted to specific niches and areas of active remodeling or high mechanical strain. However, while most healthy tissues contain little TNC, its transient expression upon cellular stress or tissue injury helps to mediate repair and restore homeostasis. Persistent expression of TNC is associated with chronic inflammation, fibrosis, and cancer, where methods for its detection are emerging as a reliable means to predict disease onset, prognosis, and response to treatment. Because studying the expression of this large matrix molecule is not always straightforward, here we describe basic techniques to examine tissue levels of TNC mRNA and protein. We also describe methods for purifying recombinant TNC, knocking down its expression, and creating cell-derived matrices with or without TNC within.
Mots clés
Cell-derived matrix, ELISA, Immunofluorescence, Immunohistochemistry, Purification, Splicing, Tenascin-C tissue expression, Western blot, qPCR, shRNA-mediated knockdown
Référence
Methods Cell Biol.. 2018 ;143:371-400