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. 2017 Nov 14;16:197. doi: 10.1186/s12934-017-0801-y

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© The Author(s) 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 2 — Preparation of Sak and its PEGylation mutants. a The Sak-cys protein (Sak-C79G) was purified by Ni²⁺-NTA affinity chromatography. Lane 1: protein marker; lane 2: pellet; lane 3: supernatant; lane 4: flow through; lane 5: elution with washing buffer; lane 6: elution with elution buffer. b Size exclusion gel filtration chromatography of Sak using the Superdex 200 column. c Figures with Coomassie blue staining (upper part) and identification of iodine staining (lower part) are combined together. Lane 1: protein marker; lane 2: reaction mixtures; lane 3: elution of peak 1; lane4: elution of peak 2; lane 5: elution of peak 3. d SDS-PAGE analysis of the PEG-Sak proteins. Lane 1: protein marker; lane 2: PEG-Sak-C79G; lane 3: PEG-Sak-C82L; lane 4: PEG-Sak-C84K; lane 5: PEG-Sak-C97A; lane 6: PEG-Sak-C104R