title: [Short gel method for pretreatment of protein samples with high concentration of detergent].
abstract: In proteomic research, to improve protein solubility of membrane proteins and nuclear proteins, buffers containing high concentration of detergent, such as 4% SDS, were widely used. However, high concentration of detergent might severely interfere with the downstream proteomic analysis, including protein quantitation and trypsin digestion. To improve the proteomic compatibility of buffers with high concentration of detergent, we used short gel method to pretreat buffers containing detergent. Protein samples were first separated by a short (2-2.5 mm) SDS-PAGE electrophoresis, and proteins were quantitated by comparing with bovine serum albumin standards via optical density analysis. The gel was then cut and peptides were recovered using in-gel digestion. The quantitative linearity range of this method was 1 to 8 μg. The quantitation was accurate and reproducible. After short gel analysis, recovered peptides generated high mass spectrometry signals. In conclusion, short gel method eliminated the interference of high concentration detergent in the proteomics analysis, and it was suitable for protein samples' pretreatment, and was worth to apply in proteomic research.
authors.forename: Shouzhi
authors.initials: S
authors.lastname: Ma
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Linhui
authors.initials: L
authors.lastname: Zhai
authors.forename: Yulin
authors.initials: Y
authors.lastname: Sun
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
authors.forename: Xiaohang
authors.initials: X
authors.lastname: Zhao
mesh_terms: D003902:Detergents / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000737:chemistry; D009687:Nuclear Proteins / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D014357:Trypsin
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D009687:Nuclear Proteins; D011506:Proteins; D014357:Trypsin