title: [Application of membrane protein-based two-dimensional electrophoresis in chondrocyte- related investigations].
abstract: OBJECTIVE
  To explore the feasibility of membrane protein-based two-dimensional electrophoresis (2-DE) in the investigations of chondrocyte-related diseases and its complementarity with total protein-based 2-DE.
  
  
  METHODS
  Knee cartilage samples were obtained to isolate the chondrocytes with type II collagenase/hyaluronidase digestion. The membrane proteins and total proteins were extracted and loaded separately onto PH3.0-10.0 non-linear gel strip for 2-DE analysis. The qualities of membrane protein-based 2-DE gels were evaluated, and the difference between the distribution profiles of the membrane protein spots and that of the total protein were observed and their complementarities were evaluated.
  
  
  RESULTS
  Membrane protein-based 2-DE generated high-quality gel images, and on each gel 412.3±13.5 protein spots were identified. These spots were distributed in the region of isoelectric point pH 5.0-9.0. In the gel images generated by the total proteins, 564.3±5.9 protein spots were identified in each image, and the spots were distributed in the region of isoelectric point pH 3.0-7.0.
  
  
  CONCLUSIONS
  Membrane protein-based 2-DE of the chondrocytes can generate high-quality gel images, and the isoelectric distribution of the protein spots is complementary to that of total protein, which provides valuable information for chondrocyte-related diseases.
authors.affiliation: Department of Orthopedics, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xiao
authors.forename: Zhan-jun
authors.initials: ZJ
authors.lastname: Shi
authors.forename: Zhan-dong
authors.initials: ZD
authors.lastname: Zhao
authors.forename: Ya-peng
authors.initials: YP
authors.lastname: Zhou
authors.forename: Tian
authors.initials: T
authors.lastname: Lan
mesh_terms: D000328:Adult; D019902:Chondrocytes / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D006801:Humans; D007526:Isoelectric Point; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification*; D008875:Middle Aged
chemical_list: D008565:Membrane Proteins