Analysis of proteomic components of sera from patients with uremia by two dimensional electrophoresis and matrix assisted laser desorption/ ionization time of flight mass spectrometry.
The different sera proteomic components between uremia patients and normal subjects were studied through two-dimensional gel electrophoresis technique. Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE), silver staining, ImageMaster 2D 5.0 analysis software, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS) and IPI human database searching were used to separate and identify the proteome of the sera from the patients with uremia. The results showed that satisfactory 2DE patterns of the serum proteins were obtained. Twenty-six protein spots showed significant difference in quantity in uremia patients, and 20 protein spots were identified by MALDI-TOF-TOF-MS. It was concluded that good reproducibility could be obtained by applying immobilized pH gradient 2DE to separate and identify the proteome in serum, which provided the foundation for the further study on uremia toxins pertaining to protein.
Wang
Jianqing
J
Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Dai
Yong
Y
Deng
Anguo
A
Liu
Jianjun
J
D000328:Adult; D001797:Blood Protein Electrophoresis; D001798:Blood Proteins / Q000032:analysis*; D016022:Case-Control Studies; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014511:Uremia / Q000097:blood*
D001798:Blood Proteins; D020543:Proteome