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. 2019 Jun 12;10:1353. doi: 10.3389/fimmu.2019.01353

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Copyright © 2019 Myngbay, Bexeitov, Adilbayeva, Assylbekov, Yevstratenko, Aitzhanova, Matkarimov, Adarichev and Kunz.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Detection and stability of plasma CTHRC1. (A) The specificity of the immunodetection was tested using 25 ng rhCTHRC1 spiked into the plasma (3 μL) of RA patients. Protein ladder bands (kD) are shown. (B) The resistance of the protein to proteolysis was tested with rhCTHRC1 spiked into synovial fluid (SF) or the plasma of RA patients. Final concentration of SF or plasma in the test was 12%, final amount of rhCTHRC1 loaded per lane was 15 ng. Lane 1 shows 50 ng of recombinant rhCTHRC1 protein as a reference. The Vli55 antibody was used for CTHRC1 immunodetection followed by appropriate secondary antibodies. As a positive control for digestion, incubation with trypsin was performed (lane 9). Lanes 10, 11: SF was heated for 30 min at 65°C (time, min) and then incubated with rhCTHRC1. Lanes 12, 13: SF was pre-heated for 30 min at 37°C.