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Fluorescence Approaches to Growing Macromolecule CrystalsTrace fluorescent labeling, typically < 1%, can be a powerful aid in macromolecule crystallization. Precipitation concentrates a solute, and crystals are the most densely packed solid form. The more densely packed the fluorescing material, the more brightly the emission from it, and thus fluorescence intensity of a solid phase is a good indication of whether one has crystals or not. The more brightly fluorescing crystalline phase is easily distinguishable, even when embedded in an amorphous precipitate. This approach conveys several distinct advantages: one can see what the protein is doing in response to the imposed conditions, and distinguishing between amorphous and microcrystalline precipitated phases are considerably simpler. The higher fluorescence intensity of the crystalline phase led us to test if we could derive crystallization conditions from screen outcomes which had no obvious crystalline material, but simply "bright spots" in the precipitated phase. Preliminary results show that the presence of these bright spots, not observable under white light, is indeed a good indicator of potential crystallization conditions.
Document ID
20070002159
Acquisition Source
Marshall Space Flight Center
Document Type
Preprint (Draft being sent to journal)
Authors
Pusey, Marc
(NASA Marshall Space Flight Center Huntsville, AL, United States)
Forsythe, Elizabeth
(Nektar Therapeutics Huntsville, AL, United States)
Achari, Aniruddha
(Raytheon Information Technology and Scientific Services Huntsville, AL, United States)
Date Acquired
August 24, 2013
Publication Date
January 1, 2006
Subject Category
Chemistry And Materials (General)
Funding Number(s)
CONTRACT_GRANT: GM071581-01
Distribution Limits
Public
Copyright
Other

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