Some practical guidelines for UV imaging in the protein crystallization laboratory
نویسندگان
چکیده
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.
منابع مشابه
Evaluating the efficacy of tryptophan fluorescence and absorbance as a selection tool for identifying protein crystals
The environment of individual tryptophans in known protein structures and the effectiveness of four commercial robotic UV microscopes to illuminate tryptophan-containing protein crystals by either tryptophan fluorescence (epi-illumination) or absorbance (transmission) are evaluated. In agreement with other studies, tryptophan residues are found on average to be largely buried in protein structu...
متن کاملبازنگری راهنمای اخلاقی کار با حیوانات آزمایشگاهی
Although we have diverse methods in laboratory studies, such as cellular and molecular sciences, understanding many issues related to human health requires research on animals. An ethical duty of every researcher is to respect animal rights. In our country, educational and research activities are carried out frequently on animals. Therefore, guidelines for ethical use of animals in Iran were de...
متن کاملAppropriateness of physicians’ lumbosacral MRI requests in private and public centers in Tehran, Iran
Background: Back pain is a common patients’ complaint, and its etiology is important because of different potential treatment approaches (based on causes). For a better diagnosis, magnetic resonance imaging (MRI) is widely used in clinical settings that may result in inappropriate requests. This study aims to evaluate the appropriateness of the lumbosacral MRI requests in patients with ba...
متن کاملA procedure for setting up high-throughput nanolitre crystallization experiments. Crystallization workflow for initial screening, automated storage, imaging and optimization.
Crystallization trials at the Division of Structural Biology in Oxford are now almost exclusively carried out using a high-throughput workflow implemented in the Oxford Protein Production Facility. Initial crystallization screening is based on nanolitre-scale sitting-drop vapour-diffusion experiments (typically 100 nl of protein plus 100 nl of reservoir solution per droplet) which use standard ...
متن کاملAssessment methods for laboratory skills
Today rapid and progressive improvements in clinical laboratory science are provided in terms of diagnostic tests technology. These changes call for educated experts with sufficient practical skills who can use new technologies and modern equipment. In other words, the employment of experienced and skilled laboratory technicians and technologists, as the main assets of each laboratory, is a com...
متن کامل