title: Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis.
abstract: OBJECTIVE
  Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established.
  
  
  METHODS
  The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimensional electrophoresis (2DE) profiling quality, including number of protein spots and spot distribution.
  
  
  RESULTS
  The total protein of bile fluid in benign biliary disorders was 0.797 ± 0.465 μg/μL. The sample preparation method using acetone precipitation first followed by 2D Clean-Up kit protein extraction resulted in better quality of 2DE gel images in terms of resolution as compared with other sample preparation methods. Using this protocol, we obtained approximately 558 protein spots on the gel images and with better protein spots presentation of haptoglobin, serum albumin, serotransferrin, and transthyretin.
  
  
  CONCLUSIONS
  Protein samples of bile prepared using acetone precipitation followed by 2D Clean-Up kit exhibited high protein resolution and significant protein profile. This optimized protein preparation protocol can effectively concentrate bile proteins, remove abundant proteins and debris, and yield clear presentation of nonabundant proteins and its isoforms on 2-dimensional electrophoresis gel images.
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Linkou Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Hao-Tsai
authors.initials: HT
authors.lastname: Cheng
authors.affiliation: Department of Gastroenterology and Hepatology, Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan.
authors.forename: Sen-Yung
authors.identifier: 0000-0002-1723-7261
authors.initials: SY
authors.lastname: Hsieh
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Linkou Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Chang-Mu
authors.initials: CM
authors.lastname: Sung
authors.affiliation: Department of Craniofacial Orthodontics, Chang Gung Memorial Hospital, Taipei, Taiwan; Graduate Institute of Craniofacial and Oral Science, Chang Gung University, Taoyuan, Taiwan; Craniofacial Research Center, Chang Gung Memorial Hospital, Linkou, Taiwan.
authors.forename: Betty Chien-Jung
authors.initials: BC
authors.lastname: Pai
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Linkou Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan.
authors.forename: Nai-Jen
authors.initials: NJ
authors.lastname: Liu
authors.affiliation: Department of Physical Medicine & Rehabilitation, Chang Gung Memorial Hospital at Linkou and College of Medicine, Chang Gung University, Kwei-Shan, Taoyuan, Taiwan.
authors.forename: Carl  P C
authors.initials: C
authors.lastname: Chen
mesh_terms: D000096:Acetone / Q000737:chemistry; D001646:Bile / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D014176:Protein Biosynthesis / Q000235:genetics; D020543:Proteome / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
chemical_list: D020543:Proteome; D000096:Acetone