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. 2014 Jan 23;9(1):e87521. doi: 10.1371/journal.pone.0087521

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© 2014 Dulle et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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The oligomeric distribution of recombinant wild type (WT) Hsp104 (blue, A & B) and (A) Hsp104-V426I (red), Hsp104-K480C (yellow), and Hsp104-Y507D (green), or (B) Hsp104-V426C (orange) and Hsp104-D434A (purple), was analyzed by ultracentrifugation through a linear glycerol gradient in the presence of 5 mM ATP. Equal fractions from the gradients were collected and analyzed by western blot with an anti-Hsp104 antibody. The amount of Hsp104 in each fraction was quantified by ImageJ and graphed as a fraction of the total Hsp104. The gradients were repeated twice with recombinant protein from two separate recombinant protein purification preparations.