Insoluble protein purification with sarkosyl: facts and precautions

Ben Chisnall, Courtney Johnson, Yavuz Kulaberoglu, Yu Wai Chen

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Citations (Scopus)

Abstract

When eukaryotic proteins are overexpressed in Escherichia coli hosts, they often form inclusion bodies. Natively folded proteins can be extracted from inclusion bodies using mild detergents such as sarkosyl. One common problem is the sequestration of nucleic acid contaminants with the protein of interest. Here we describe methods for monitoring the presence of co-precipitated nucleic acids, and their removal. These procedures are simple to implement and can be easily adapted to a high-throughput format. While sarkosyl is a common chemical, some information such as its UV absorption spectrum and micellar size are absent in the literature or poorly referenced. We review and summarize the properties that are the most relevant to structural biology.
Original languageEnglish
Title of host publicationStructural Genomics
Subtitle of host publicationGeneral Applications
EditorsYu Wai Chen
Place of PublicationNew York
PublisherHumana Press
Pages179-186
Number of pages8
VolumeN/A
EditionN/A
ISBN (Print)9781627036900
DOIs
Publication statusPublished - 30 Nov 2013

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume1091
ISSN (Print)1064-3745

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