Products/Services Used | Details | Operation |
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PCR Cloning and Subcloning> | ... LCs. BoNT/E LC was purchased from Genscript (USA) and cloned to modify pET28a vector. The cloned BoNT/E LC was purified using Ni-NTA (Qiagen) affinity chromatography and a Superdex S200 column (GE Healthcare). ... | Get A Quote |
We report a simple label-free electrochemical method of detecting low concentrations of botulinum neurotoxin type E light chain (BoNT/E LC) based on its peptide cleavage activity. Dual-mode cyclic voltammetry was employed to observe changes in the redox signal of ferri-/ferro-cyanide on interdigitated microelectrodes, whose surfaces were covered by peptides designed from synaptosomal-associated protein 25 to be cleaved by BoNT/E LC. With the introduction of BoNT/E LC, the redox signal showed a time-dependent increase due to cleavage of the immobilized peptide molecules. In addition to the increased redox signal intensity, its time-dependence can be considered as a strong evidence of BoNT/E sensing, since the ti... More