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Latest methods of fluorescence‐based protein crystal identification
Author(s) -
Meyer Arne,
Betzel Christian,
Pusey Marc
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15000114
Subject(s) - fluorescence , protein crystallization , covalent bond , crystallization , crystal (programming language) , identification (biology) , chemistry , fluorescent protein , materials science , biophysics , nanotechnology , computer science , optics , biochemistry , biology , green fluorescent protein , physics , organic chemistry , botany , gene , programming language
Successful protein crystallization screening experiments are dependent upon the experimenter being able to identify positive outcomes. The introduction of fluorescence techniques has brought a powerful and versatile tool to the aid of the crystal grower. Trace fluorescent labeling, in which a fluorescent probe is covalently bound to a subpopulation (<0.5%) of the protein, enables the use of visible fluorescence. Alternatively, one can avoid covalent modification and use UV fluorescence, exploiting the intrinsic fluorescent amino acids present in most proteins. By the use of these techniques, crystals that had previously been obscured in the crystallization drop can readily be identified and distinguished from amorphous precipitate or salt crystals. Additionally, lead conditions that may not have been obvious as such under white‐light illumination can be identified. In all cases review of the screening plate is considerably accelerated, as the eye can quickly note objects of increased intensity.