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

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