title: Simple sodium dodecyl sulfate-assisted sample preparation method for LC-MS-based proteomics applications.
abstract: Sodium dodecyl sulfate (SDS) is one of the most popular laboratory reagents used for biological sample extraction; however, the presence of this reagent in samples challenges LC-MS-based proteomics analyses because it can interfere with reversed-phase LC separations and electrospray ionization. This study reports a simple SDS-assisted proteomics sample preparation method facilitated by a novel peptide-level SDS removal step. In an initial demonstration, SDS was effectively (>99.9%) removed from peptide samples through ion substitution-mediated DS(-) precipitation using potassium chloride (KCl), and excellent peptide recovery (>95%) was observed for <20 μg of peptides. Further experiments demonstrated the compatibility of this protocol with LC-MS/MS analyses. The resulting proteome coverage obtained for both mammalian tissues and bacterial samples was comparable to or better than that obtained for the same sample types prepared using standard proteomics preparation methods and analyzed using LC-MS/MS. These results suggest the SDS-assisted protocol is a practical, simple, and broadly applicable proteomics sample processing method, which can be particularly useful when dealing with samples difficult to solubilize by other methods.
authors.affiliation: Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, United States.
authors.forename: Jian-Ying
authors.initials: JY
authors.lastname: Zhou
authors.forename: Geoffrey P
authors.initials: GP
authors.lastname: Dann
authors.forename: Tujin
authors.initials: T
authors.lastname: Shi
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Gao
authors.forename: Dian
authors.initials: D
authors.lastname: Su
authors.forename: Carrie D
authors.initials: CD
authors.lastname: Nicora
authors.forename: Anil K
authors.initials: AK
authors.lastname: Shukla
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Moore
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Qian
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods; D005590:Fractional Precipitation / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis* / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D020592:Shewanella / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry / Q000302:isolation & purification*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D020543:Proteome; D012967:Sodium Dodecyl Sulfate