title: Improved 2-nitrobenzenesulfenyl method: optimization of the protocol and improved enrichment for labeled peptides.
abstract: We have developed the NBS (2-nitrobenzenesulfenyl) method, a quantitative proteome analysis method utilizing stable isotope labeling followed by mass spectrometry. The potential of this method was reported previously, and the procedure has now been further optimized. Here, we describe a procedure utilizing urea or guanidine hydrochloride as a protein denaturant, in conjunction with an improved chromatographic enrichment method for the NBS-labeled peptides using a phenyl resin column. By using this new protocol, both sample loss throughout the protocol and the elution of unwanted unlabeled peptides can be minimized, improving the efficiency of the analysis significantly.
authors.affiliation: Life Science Laboratory, Shimadzu Corporation, Kyoto 604-8511, Japan.
authors.forename: Ei-Ichi
authors.initials: E
authors.lastname: Matsuo
authors.forename: Chikako
authors.initials: C
authors.lastname: Toda
authors.forename: Makoto
authors.initials: M
authors.lastname: Watanabe
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Iida
authors.forename: Taro
authors.initials: T
authors.lastname: Masuda
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Minohata
authors.forename: Eiji
authors.initials: E
authors.lastname: Ando
authors.forename: Susumu
authors.initials: S
authors.lastname: Tsunasawa
authors.forename: Osamu
authors.initials: O
authors.lastname: Nishimura
mesh_terms: D000818:Animals; D002645:Chickens; D019791:Guanidine; D051379:Mice; D009578:Nitrobenzenes / Q000737:chemistry*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry*; D011489:Protein Denaturation; D040901:Proteomics / Q000379:methods*; D020539:Sequence Analysis, Protein; D014508:Urea
chemical_list: D009578:Nitrobenzenes; D010455:Peptides; C002732:2-nitrobenzenesulfenyl chloride; D014508:Urea; D019791:Guanidine