title: Proteome analysis of heart biopsies using a TRIzol-based protein extraction.
abstract: BACKGROUND
  When dealing with rare samples of which only minute amounts are available, e.g. human heart tissue, simultaneous extraction of DNA, RNA, and proteins from the same sample is crucial for a comprehensive analysis on the physiological or pathological state of such precious tissue. In this study we provethe efficacy of a modified TRIzol protocol to extract proteins from samples of small size, such as endomyocardial biopsies (EMBs).
  
  
  METHODS
  Initially, we compared TRIzol protein extraction efficacy to urea/thiourea extraction from total murine left ventricles and then small amounts of left and right murine ventricles. Finally, we applied the modified TRIzol protocol to the proteomic profiling of EMBs from human left and right ventricles.
  
  
  RESULTS
  Analysis of the proteins extracted from mouse and human samples revealed sufficient protein amount for downstream applications. Thus, LC-tandem mass spectrometry permitted highly sensitive protein identifications and comparable protein patterns and coverage of cellular components as a standard extraction protocol. 2D gel-based analysis confirmed the high quality and reproducibility of the TRIzol derived protein extracts.
  
  
  CONCLUSIONS
  Our results prove the utility of the modified TRIzol protocol for proteomics analyses involving minute amounts of precious samples.
authors.affiliation: Department of Pharmacology, Physiology and Pathophysiology, Faculty of Pharmacy, University of Medicine and Pharmacy "Iuliu Hatieganu", Cluj-Napoca, Romania; Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: pop.cristina@umfcluj.ro.
authors.forename: Cristina
authors.initials: C
authors.lastname: Pop
authors.affiliation: Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: amelings@uni-greifswald.de.
authors.forename: Sabine
authors.initials: S
authors.lastname: Ameling
authors.affiliation: Department of Internal Medicine B, University Medicine Greifswald, Germany. Electronic address: empen@uni-greifswald.de.
authors.forename: Klaus
authors.initials: K
authors.lastname: Empen
authors.affiliation: Department of Internal Medicine B, University Medicine Greifswald, Germany. Electronic address: ruedebu@uni-greifswald.de.
authors.forename: Julia
authors.initials: J
authors.lastname: Rüdebusch
authors.affiliation: Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: dhoplevm@uni-greifswald.de.
authors.forename: Vishnu Mukund
authors.initials: VM
authors.lastname: Dhople
authors.affiliation: Department of Internal Medicine B, University Medicine Greifswald, Germany; DZHK (German Center for Cardiovascular Research, Partner Site Greifswald), Germany. Electronic address: felix@uni-greifswald.de.
authors.forename: Stephan B
authors.initials: SB
authors.lastname: Felix
authors.affiliation: Department of Toxicology, Faculty of Pharmacy, University of Medicine Pharmacy "Iuliu Hatieganu", Cluj-Napoca, Romania. Electronic address: floghin@umfcluj.ro.
authors.forename: Felicia
authors.initials: F
authors.lastname: Loghin
authors.affiliation: Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany; DZHK (German Center for Cardiovascular Research, Partner Site Greifswald), Germany. Electronic address: voelker@uni-greifswald.de.
authors.forename: Uwe
authors.initials: U
authors.lastname: Völker
authors.affiliation: Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany; DZHK (German Center for Cardiovascular Research, Partner Site Greifswald), Germany. Electronic address: hammer@uni-greifswald.de.
authors.forename: Elke
authors.initials: E
authors.lastname: Hammer
mesh_terms: D000818:Animals; D001706:Biopsy; D002853:Chromatography, Liquid; D006146:Guanidines / Q000737:chemistry; D006352:Heart Ventricles / Q000737:chemistry*; D006801:Humans; D059627:Liquid Phase Microextraction / Q000379:methods*; D051379:Mice; D058977:Molecular Sequence Annotation; D009206:Myocardium / Q000737:chemistry*; D010636:Phenols / Q000737:chemistry; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry; D013890:Thiourea / Q000737:chemistry; D014508:Urea / Q000737:chemistry
chemical_list: D006146:Guanidines; D010636:Phenols; D020543:Proteome; C411644:trizol; D014508:Urea; D013890:Thiourea
keywords: 2D PAGE; Heart biopsies; LC–MS/MS; Protein extraction; Proteomics; TRIzol