Large-scale amino-acid analysis for proteome studies.
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.
Yan
J X
JX
Macquarie University Centre for Analytical Biotechnology, Macquarie University, Sydney, NSW, Australia.
Wilkins
M R
MR
Ou
K
K
Gooley
A A
AA
Williams
K L
KL
Sanchez
J C
JC
Golaz
O
O
Pasquali
C
C
Hochstrasser
D F
DF
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
D000596:Amino Acids; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers