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Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Feb 1;69(Pt 2):201-8. doi: 10.1107/S1744309112048634. Epub 2013 Jan 26.

Some practical guidelines for UV imaging in the protein crystallization laboratory.

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1
CSIRO Materials, Science and Engineering, 343 Royal Parade, Parkville, VIC 3052, Australia.

Abstract

High-throughput imaging of protein crystallization experiments with ultraviolet (UV) light has recently become commercially available and can enable crystallographers to differentiate between crystals of protein and those of salt, as the visualization of protein crystals is based on intrinsic tryptophan fluorescence. Unfortunately, UV imaging is not a panacea, as some protein crystals will not fluoresce under UV excitation and some salt crystals are UV-fluorescently active. As a new technology, there is little experience within the general community on how to use this technology effectively and what caveats to look out for. Here, an attempt is made to identify some of the common problems that may arise using UV-imaging technology by examining test proteins, common crystallization reagents and a range of proteins by assessing their UV-Vis absorbance spectra. Some pointers are offered as to which systems may not be appropriate for this methodology.

KEYWORDS:

UV imaging

PMID:
23385768
PMCID:
PMC3564629
DOI:
10.1107/S1744309112048634
[Indexed for MEDLINE]
Free PMC Article
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