Some practical guidelines for UV imaging in the protein crystallization laboratory
Author(s) -
Desbois Sebastien,
Seabrook Shane A.,
Newman Janet
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112048634
Subject(s) - protein crystallization , crystallization , ultraviolet , fluorescence , absorbance , nanotechnology , panacea (medicine) , materials science , salt (chemistry) , chemistry , computer science , optics , optoelectronics , chromatography , physics , organic chemistry , pathology , alternative medicine , medicine
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.
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