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. 2019 Dec 17;9:19242. doi: 10.1038/s41598-019-55805-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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ELISA and SDS-PAGE analysis under thermal stress. (a,c,e) ELISA of the antigen-binding activity of 3D8 antibodies. Purified 3D8-derived antibodies were heated under the specified conditions, placed in wells coated with plasmid DNA antigen (pUC19), and bound 3D8 antibodies were detected with AP-conjugated antibodies specific for mouse IgG/Fc (a), human IgG/Fc (b), or chicken IgY/υ chain (c). Data are presented as mean ± SD (n = 3). (b,d,f) SDS-PAGE analysis of antibody integrity. Purified 3D8 antibodies were heated under the specified conditions then subjected to SDS-PAGE under reducing conditions using a 12% polyacrylamide gel, followed by staining with Coomassie Blue.