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. 2019 Aug 22;9:12269. doi: 10.1038/s41598-019-48555-w

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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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The B12-DFO conjugate was first incubated with ⁸⁹Zr at neutral pH at room temperature for 15 min according to previously published protocols²⁸. After confirmation of binding through iTLC, [⁸⁹Zr]-B12-DFO was incubated with a slight excess of apo-IF (indicated in red) for 30 min then purified with a 30 kDa spin filter (GE Vivaspin).