| Literature DB >> 25950961 |
Chelsy Prince1, Zongchao Jia2.
Abstract
Membrane protein crystallography has the potential to greatly aid our understanding of membrane protein biology. Yet, membrane protein crystals remain challenging to produce. Although robust methods for the expression and purification of membrane proteins continue to be developed, the detergent component of membrane protein samples is equally important to crystallization efforts. This chapter describes the development of three colorimetric assays for the quantitation of detergent in membrane protein samples and provides detailed protocols. All of these techniques use small sample volumes and have potential applications in crystallography. The application of these techniques in crystallization prescreening, detergent concentration modification, and detergent exchange experiments is demonstrated. It has been observed that the concentration of detergent in a membrane protein sample can be just as important as the protein concentration when attempting to reproduce crystallization lead conditions.Keywords: Analytical methods; Colorimetry; Crystallization; Detergent; Membrane protein; Quantification
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Year: 2015 PMID: 25950961 DOI: 10.1016/bs.mie.2014.12.002
Source DB: PubMed Journal: Methods Enzymol ISSN: 0076-6879 Impact factor: 1.600