Hydroxylamine Chemical Digestion for Insoluble Extracellular Matrix Characterization.

ABSTRACT: The extracellular matrix (ECM) is readily enriched by decellularizing tissues with nondenaturing detergents to solubilize and deplete the vast majority of cellular components. This approach has been used extensively to generate ECM scaffolds for regenerative medicine technologies and in 3D cell culture to model how the ECM contributes to disease progression. A highly enriched ECM fraction can then be generated using a strong chaotrope buffer that is compatible with downstream bottom-up proteomic analysis or 3D cell culture experiments after extensive dialysis. With most tissues, an insoluble pellet remains after chaotrope extraction that is rich in structural ECM components. Previously, we showed that this understudied fraction represented approximately 80% of total fibrillar collagen from

SUBMITTER: Barrett AS 

PROVIDER: S-EPMC5802359 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature