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. 2014 Nov 14;9(11):e112490. doi: 10.1371/journal.pone.0112490

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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(A) The affinity-purified rabbit anti-PP antibody, whole antisera (unpurified anti-PP antisera), and column flow-through solution were titrated using ELISA through serial dilutions ranging from 1∶1,000 to 1∶64,000. Each value represents the mean of triplicate determinations. (B) To examine the specificity of this rabbit anti-PP antibody, the dentin extract was dot-blotted onto a nitrocellulose membrane and PP was detected with the rabbit anti-PP antibody (1∶2,500). (C) The rPP protein (500 ng) appeared as a single band on SDS-PAGE, as observed with Stains-All staining, and the molecular size of this single band was concomitant with the protein band identified by anti-PP. The rC-DMP-1 protein (1 µg) appeared as a single band on SDS-PAGE, as observed using Stains-All and Coomassie brilliant (CBB) staining, which confirmed the purities of these rPP and rC-DMP-1 proteins for further experimental use. Lane M, molecular weight marker.