Isolation of proteins by heat-induced extraction from formalin-fixed, paraffin-embedded tissue and preparation of tryptic peptides for mass spectrometric analysis.
This unit describes a method of isolating of proteins from formalin-fixed and paraffin-embedded (FFPE) tissue for mass spectrometry analysis. Heat-induced antigen retrieval is the basis of the protein extraction strategy presented in this protocol. This protocol may be used to identify nuclear, cytosolic, and membrane proteins from FFPE tissues extracted from tissue blocks or slides.
Tian
Yuan
Y
Johns Hopkins University, Baltimore, Maryland, USA.
Zhang
Hui
H
D000818:Animals; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry*; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis; D051381:Rats; D016707:Tissue Fixation / Q000379:methods*
D005404:Fixatives; D010455:Peptides; D011506:Proteins; D020543:Proteome; D005557:Formaldehyde