January 1, 2013
Journal Of The Serbian Chemical Society
Product: Botulinum Neurotoxin Type B Light Chain, Recombinant
In vitro BoNT LC metalloprotease activity:
Determination of BoNT/A LC percent inhibition by SMNPIs was performed as previously described. For BoNT/B LC percent inhibition, the HTS-assay utilizes a commercially available fluorogenic substrate from List Biological Laboratories (Campbell, CA). Briefly, a multichannel pipette was used to add fluorescent substrate (final concentration 20 M) and small molecules (final concentration 20 M) to a 96 well microplate. The reactions were initiated by adding recombinant BoNT/B LC (final concentration 40 nM) to each well. All reactions were conducted in a buffered solution consisting of 40 mM Hepes pH 7.2, 1 mM DTT and 100 M ZnCl2. The change in fluorescence intensity over time was continuously monitored in each well using a 96 well plate fluorimeter (Saphire 2, Tecan, Mnnedorf, Switzerland). The assays were run at 37 C, quenched by the addition of TFA, and analyzed using reverse-phase HPLC. Potential SMNPIs of the BoNT B LC were identified by comparing the reaction velocities (change in fluorescence intensity over time) of samples assayed in the presence of compounds versus control samples.
Author did not specify which List Biological Laboratories product was utilized. Refer to the follwing List Labs link for Botulinum products available: https://listlabs.com/product-information/botulinum-toxins/