title: Large-scale amino-acid analysis for proteome studies.
abstract: Amino-acid analysis is a relatively new method for identification of proteins separated by two-dimensional gel electrophoresis and blotted onto polyvinylidene difluoride (PVDF) membranes. This article describes modified amino-acid analysis methods for this purpose. Streamlined sample handling is a key feature of the process. To minimise sample manipulation, a single vial is used for hydrolysis and the protein hydrolysate on PVDF membrane is extracted by a one-step procedure. The hydrolysate should not be stored for long periods before analysis. Applications of the technique are presented to demonstrate the identification procedure. This approach is the most cost-effective and time-effective first step in mass protein screening for a large-scale proteome project.
authors.affiliation: Macquarie University Centre for Analytical Biotechnology, Macquarie University, Sydney, NSW, Australia.
authors.forename: J X
authors.initials: JX
authors.lastname: Yan
authors.forename: M R
authors.initials: MR
authors.lastname: Wilkins
authors.forename: K
authors.initials: K
authors.lastname: Ou
authors.forename: A A
authors.initials: AA
authors.lastname: Gooley
authors.forename: K L
authors.initials: KL
authors.lastname: Williams
authors.forename: J C
authors.initials: JC
authors.lastname: Sanchez
authors.forename: O
authors.initials: O
authors.lastname: Golaz
authors.forename: C
authors.initials: C
authors.lastname: Pasquali
authors.forename: D F
authors.initials: DF
authors.lastname: Hochstrasser
mesh_terms: D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D002417:Cattle; D015698:Genomic Library; D006801:Humans; D006868:Hydrolysis; D011145:Polyvinyls; D011506:Proteins / Q000737:chemistry*; D005465:Fluorocarbon Polymers
chemical_list: D000596:Amino Acids; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers