title: Establishment of the optimum two-dimensional electrophoresis system of ovine ovarian tissue.
abstract: Lambing performance of sheep is the most important economic trait and is regarded as a critic factoring affecting the productivity in sheep industry. Ovary plays the most roles in lambing trait. To establish the optimum two-dimensional electrophoresis system (2-DE) of ovine ovarian tissue, the common protein extraction methods of animal tissue (trichloroacetic acid/acetone precipitation and direct schizolysis methods) were used to extract ovine ovarian protein, and 17-cm nonlinear immobilized PH 3-10 gradient strips were used for 2-DE. The sample handling, loading quantity of the protein sample, and isoelectric focusing (IEF) steps were manipulated and optimized in this study. The results indicate that the direct schizolysis III method, a 200-μg loading quantity of the protein sample, and IEF steps II (20°C active hydration, 14 h→500 V, 1 h→1000 V 1 h→1000-9000 V, 6 h→80,000 VH→500 V 24 h) are optimal for 2-DE analysis of ovine ovarian tissue. Therefore, ovine ovarian tissue proteomics 2-DE was preliminarily established by the optimized conditions in this study; meanwhile, the conditions identified herein could provide a reference for ovarian sample preparation and 2-DE using tissues from other animals.
authors.affiliation: The Key Laboratory of Animal Genetic Breeding and Reproduction, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
authors.forename: J L
authors.initials: JL
authors.lastname: Jia
authors.affiliation: The Key Laboratory of Animal Genetic Breeding and Reproduction, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China zhangliping@gsau.edu.cn.
authors.forename: L P
authors.initials: LP
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of Animal Genetic Breeding and Reproduction, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
authors.forename: J P
authors.initials: JP
authors.lastname: Wu
authors.affiliation: The Key Laboratory of Animal Genetic Breeding and Reproduction, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.affiliation: The Key Laboratory of Animal Genetic Breeding and Reproduction, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Ding
mesh_terms: D000096:Acetone / Q000737:chemistry; D000818:Animals; D011232:Chemical Precipitation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D010053:Ovary / Q000378:metabolism*; D020543:Proteome / Q000032:analysis* / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012756:Sheep; D014238:Trichloroacetic Acid / Q000737:chemistry
chemical_list: D020543:Proteome; D000096:Acetone; D014238:Trichloroacetic Acid