title: [Comparison of three protein extraction methods for proteomic analysis of Alexandrium tamarense with two-dimensional electrophoresis].
abstract: OBJECTIVE
  In order to find the best extraction method for proteins of Alexandrium tamarense for two-dimensional electrophoresis (2-DE) analysis.
  
  
  METHODS
  Three methods for extracting proteins from A. tamarense were compared, including trihydroxymethyl aminomethane (Tris-HCl ) buffer extraction, trichloroacetic acid (TCA)/acetone precipitation and lysis buffer extraction. Alga was cultivated in normal f/2 media (control) and supplemented with algicidal substances. Proteins obtained using the best extraction method were separated with 2-DE. Protein-expression differences were identified using matrix-assisted laser desorption/ionization-time of flight ( MALDI-TOF) mass spectrometry (MS).
  
  
  RESULTS
  Among the three protein extraction methods, lysis buffer extraction shows the best detection of the number and quality of protein spots with a clear background. Then, the lysis buffer extraction method was successfully applied to profiling protein expression in algicidal substances stress conditions and 14 differential expression proteins were identified using MALDI-TOF/MS.
  
  
  CONCLUSIONS
  Lysis buffer extraction was the most effective protein extraction method for Alexandrium tamarense.
authors.affiliation: State Key Laboratory for Marine Environmental Science, Xiamen University, Xiamen 361005, China. yawei1983@126.com
authors.forename: Yawei
authors.initials: Y
authors.lastname: Yuan
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.forename: Shaojing
authors.initials: S
authors.lastname: Li
authors.forename: Guizhong
authors.initials: G
authors.lastname: Wang
authors.forename: Tianling
authors.initials: T
authors.lastname: Zheng
mesh_terms: D004141:Dinoflagellida / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000302:isolation & purification*
chemical_list: D015800:Protozoan Proteins