Large-scale amino-acid analysis for proteome studies

被引:41
作者
Yan, JX
Wilkins, MR
Ou, K
Gooley, AA
Williams, KL
Sanchez, JC
Golaz, O
Pasquali, C
Hochstrasser, DF
机构
[1] MACQUARIE UNIV, CTR ANALYT BIOTECHNOL, SYDNEY, NSW 2109, AUSTRALIA
[2] UNIV GENEVA, MED COMP CTR, CH-1121 GENEVA 14, SWITZERLAND
[3] UNIV GENEVA, CENT CLIN CHEM LAB, CH-1121 GENEVA 14, SWITZERLAND
关键词
proteomes; amino acids; proteins;
D O I
10.1016/0021-9673(95)01285-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:291 / 302
页数:12
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