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在使用对硝基苯酯生物素(生物素-ONp)进行点合成过程中对肽进行蛋白质标记和生物素化。

Protein labeling and biotinylation of peptides during spot synthesis using biotin p-nitrophenyl ester (biotin-ONp).

作者信息

Winkler Dirk F H, McGeer Patrick L

机构信息

Peptide Array Facility, Brain Research Centre, University of British Columbia, Vancouver, Canada.

出版信息

Proteomics. 2008 Mar;8(5):961-7. doi: 10.1002/pmic.200700909.

Abstract

Biotin-labeled peptides are used for numerous biochemical and microbiological applications. Due to the strong affinity of biotin to streptavidin, the detection of biotinylated molecules is very sensitive. A powerful technique for parallel synthesis and high-throughput screening of peptides is the spot synthesis. One example for the use of spot synthesis is the screening of biotinylated peptides synthesized on cellulose membranes, which is particularly favorable for the investigation of protease cleavage sites. Additionally, in combination with biotinylated protein samples, the spot technique can be used for investigations of peptide-protein and protein-protein interactions. Here, we present our results of the use biotin p-nitrophenyl ester (biotin-ONp) in spot synthesis and as a reagent for biotin-labeling of protein samples.

摘要

生物素标记的肽可用于众多生化和微生物学应用。由于生物素与链霉亲和素的强亲和力,生物素化分子的检测非常灵敏。斑点合成是一种用于肽的平行合成和高通量筛选的强大技术。斑点合成的一个应用实例是对在纤维素膜上合成的生物素化肽进行筛选,这对于研究蛋白酶切割位点特别有利。此外,结合生物素化的蛋白质样品,斑点技术可用于研究肽-蛋白质和蛋白质-蛋白质相互作用。在此,我们展示了使用对硝基苯酯生物素(生物素-ONp)进行斑点合成以及作为蛋白质样品生物素标记试剂的结果。

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