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The button test: A small scale method using microdialysis cells for assessing protein solubility at concentrations suitable for NMR

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Abstract

A simple method has been developed for screening solutionconditions to determine conditions under which a protein is soluble at thehigh concentrations typically used for NMR spectroscopy. The method employsmicrodialysis cells or ‘buttons’. The low sample volume (5 µl)required for each microdialysis button permits testing of a wide range ofsolution conditions and temperatures with high protein concentrations, usinga small amount of protein. Following precipitation of several NMR samples ofthe C-terminal core domain of human TFIIB, the microdialysis button screenfacilitated identification of conditions in which precipitation of the TFIIBcore domain was eliminated. The microdialysis button method for screeningsolution conditions is generally applicable and has been used to permitrapid identification of suitable NMR sample solution conditions for proteinsinvolved in transcription and cell adhesion.

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Authors and Affiliations

  1. Division of Molecular and Structural Biology, Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, 610 University Avenue, Toronto, ON, Canada M5G 2M9

    Stefan Bagby, Kit I. Tong, Dingjiang Liu, Jean-René Alattia & Mitsuhiko Ikura

Authors
  1. Stefan Bagby
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  2. Kit I. Tong
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  3. Dingjiang Liu
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  4. Jean-René Alattia
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  5. Mitsuhiko Ikura
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Bagby, S., Tong, K.I., Liu, D. et al. The button test: A small scale method using microdialysis cells for assessing protein solubility at concentrations suitable for NMR. J Biomol NMR 10, 279–282 (1997). https://doi.org/10.1023/A:1018359305544

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  • DOI: https://doi.org/10.1023/A:1018359305544

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