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. 2022 Dec 9;20:578. doi: 10.1186/s12967-022-03794-w

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Confirmation of AzK incorporation in Stx1B K9AzK and scFv OKT3 E129AzK by fluorophore labelling. The purified proteins scFv OKT3 (lane 1, MW_calc 30 kDa), scFv OKT3 E129AzK (lane 2, MW_calc 30 kDa), Stx1B (lane 3, MW_calc 9 kDa) and Stx1B K9AzK (lane 4, MW_calc 9 kDa) were incubated with DBCO-Cy3 as detailed in the materials and methods section and then analyzed on a 4–12% SDS gel. a`Proteins stained with Coomassie protein stain. b SDS gel irradiated at 635 nm before staining with Coomassie protein stain. The sizes of the molecular weight marker (M) bands are indicated on the left margin of the gels