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Finally, amino-ethylcarbamazimine (AEC) was used as a substrate reagent.
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Future modification of our assay to encompass other BoNT serotypes, and even other classes of toxins, should become feasible as soon as the necessary substrate reagents become available.
The biofumigant was used as an enzyme inhibitor and acetylthiocholine iodide as a substrate along with Ellman's reagent to carry out the reactions.
The bound peroxidase is reacted with TMB substrate (Reagent 6) for a predetermined time.
Proteins were then detected using ECL reagent (Amersham, USA) as a substrate.
To get the best Dicer1 knock-down, NIH/3T3 cells, among the easiest to transfect by established morpholino reagents, were selected as a substrate.
For both primary antibodies, goat anti-rabbit immunoglobulin G conjugated with horseradish peroxidase (Pierce; Product Code: 31463); 1∶10000 in blocking buffer) was used as the second step antibody, and ChemiGlow reagent (Alpha Innotech) was used as a substrate.
Substrate reagent was added to each well, followed by a stop solution (0.5 M sulfuric acid).
The ALDEFLUOR reagent is known to act as a substrate for ALDH1.
Adherent cells were directly dissolved in 50 μl Proteasome-Glo™ lysis and reagent buffer, containing Suc-LLVY-aminoluciferin as a substrate.
TMB substrate reagent set was purchased from BD Biosciences.
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