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. 2017 Nov 9;8:796. doi: 10.3389/fphar.2017.00796

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Copyright © 2017 Rust, Doran, Hart, Binz, Stickings, Sesardic, Peden and Davletov.

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) or licensor 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 of BoNT/B activity in NanoLuc-VAMP2 SiMa cells using specific BoNT/B-cleaved VAMP2 antibody. (A) The VAMP2 amino acid sequence used for generation of the rabbit polyclonal antibody is underlined. The arrow indicates the cleavage site by BoNT/B. (B) Immunoblot showing dose-dependent increase of BoNT/B-cleaved NanoLuc-VAMP2 product using the cleaved VAMP2 antibody. Engineered SiMa cells were incubated for 60 h in the presence of indicated concentrations of BoNT/B followed by SDS–PAGE and immunoblotting. (C) Graph showing quantification of dose-dependent increase in cleaved VAMP2 immunoblotting signal in response to BoNT/B with sensitivity in the low picomolar range (n = 3 ± SD).