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Analytical Chem JoVE (Open Access) Citable · DOI

Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis

DOI: 10.3791/53057-v
What you'll learn
  • ✓ Decellularize tissue samples to enrich extracellular matrix proteins
  • ✓ Digest ECM-enriched preparations into peptides for mass spectrometry
  • ✓ Analyze ECM protein composition using proteomics workflows
  • ✓ Validate decellularization efficiency by Western blot
Protocol

This protocol describes a procedure for enriching ECM proteins from tissues or tumors and deglycosylating and digesting the ECM-enriched preparations into peptides to analyze their protein composition by mass spectrometry.

Difficulty
intermediate
Total time
~2–3 days (including incubation and digestion steps)
Biosafety
BSL-1

Steps

1
Decellularize tissue to enrich ECM proteins

Remove cellular components from tissue or tumor samples using chemical or enzymatic methods to isolate and enrich extracellular matrix proteins for downstream analysis.

▶ 01:13
2
Digest ECM proteins into peptides

Deglycosylate and enzymatically digest the ECM-enriched preparation into peptides suitable for mass spectrometry analysis of protein composition.

▶ 04:27
3
Validate decellularization by Western blot

Confirm successful removal of cellular components and enrichment of ECM proteins using Western blot analysis of marker proteins.

▶ 06:11
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