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From test tube to plate: a simple procedure for the rapid preparation of microcrystallization experiments using the cubic phase method
Author(s) -
Nollert Peter
Publication year - 2002
Publication title -
journal of applied crystallography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.429
H-Index - 162
ISSN - 1600-5767
DOI - 10.1107/s0021889802012001
Subject(s) - crystallization , materials science , protein crystallization , phase (matter) , bacteriorhodopsin , diffraction , crystallography , chromatography , computer science , chemical engineering , chemistry , optics , membrane , physics , biochemistry , organic chemistry , engineering
Crystals of transmembrane proteins for X‐ray diffraction experiments may be grown either by employing mixed protein–detergent complexes, or in a matrix of liquid‐crystalline membraneous material forming a lipidic cubic phase ( in cubo ). Widespread use of the in cubo method has been severely hampered by its tediousness and the large amounts of protein required. Here a simple procedure is presented that by virtue of its simplicity and small setup size substantially reduces the preparation time as well as the amount of protein. Crystallization trials are set up in conventional multi‐well plates using a semi‐automatic dispenser‐driven microsyringe. The microprocedure is amenable to full automation and further miniaturization. Its feasibility is demonstrated by screening for new crystallization conditions for bacteriorhodopsin using volumes of ca 200 nl of lipidic cubic phase. New crystallization conditions were identified that avoid the necessity of weighing solid precipitation agents.