Abstract:

To compare different sample preparation methods for human serum 2DE. MethodsSerum samples were prepared by five different methods: acetone precipitation, trichloroacetic acid （TCA） precipitation, acetone /TCA precipitation, 2D clean up kit, and Albumin and IgG Removal Kit. Then, after the samples were run on two dimensional gel electrophoresis (2DE), 2DE gels were stained and analyzed by Image Master 5.0, and protein spots were identified by MALDITOF MS. ResultsThe total protein number of samples processed by acetone precipitation, TCA precipitation, Acetone/TCA precipitation, and 2D clean up kit was 1?802±12, 1?648±10, 1?727±13 and 1?670±14, respectively. In addition, the total protein number of samples, one of which was processed by sample preparation kits removing the albumin and immunity goblin, the other one of which was not, was 1?212±16 and 1?258±12, respectively. Conclusion2DE maps obtained from samples treated by different serum precipitation methods were different. By removing albumin and immunity goblin from the serum, low abundance proteins were increased, but some high abundance proteins were lost.

Key words:  Two dimensional gel electrophoresis; Sample preparation; Proteomics; Serum