Rapid purification of EGFP, EYFP, and ECFP with high yield and purity

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Title Rapid purification of EGFP, EYFP, and ECFP with high yield and purity
Author McRae, Shelley Rose; Brown, Chris; Bushell, Gillian Robin
Journal Name Protein Expression and Purifcation
Editor R.R. Burgess
Year Published 2005
Place of publication US
Publisher Academic Press
Abstract Most current high throughput puriWcation procedures for the green Xuorescent protein (GFP) suVer from poor yields and low purity. An improved puriWcation procedure that delivers highly pure protein (>95% homogeneity) in high yields (>70% of the initial Xuorescent protein content) has been developed. The puriWcation procedure requires only two steps: the cell lysate is heated to 60 °C for 4min in ammonium sulfate and triethylamine, followed by hydrophobic interaction chromatography using isopropanol during the elution phase. The resulting pure product exhibits the same Xuorescence proWle as the crude sample. This procedure has been demonstrated on three commercial variants of GFP from Aequorea victoria, enhanced green, enhanced yellow, and enhanced cyan Xuorescent protein (Becton–Dickinson). The yield and purity of material are superior to other recently described methods.
Peer Reviewed Yes
Published Yes
Publisher URI http://www.elsevier.com/wps/find/journaldescription.cws_home/622935/description#description
Alternative URI http://dx.doi.org/10.1016/j.pep.2004.12.030
Copyright Statement Copyright 2005 Elsevier : Reproduced in accordance with the copyright policy of the publisher : This journal is available online - use hypertext links.
Volume 41
Page from 121
Page to 127
ISSN 1046-5928
Date Accessioned 2006-01-10
Language en_AU
Research Centre Eskitis Institute for Drug Discovery; Queensland Micro and Nanotechnology Centre
Faculty Faculty of Science
Subject PRE2009-Biotechnology
URI http://hdl.handle.net/10072/4975
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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