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To enhance sensitivity, reaction buffers were supplemented by the addition of sodium 3,5-dichloro-2-hydroxy-benzenesulfonate.
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As described in Section 4.3, the BoNTAe inhibition assay buffer was supplemented by 25 µM ZnCl2, whereas the BoNTBe inhibition assay buffer contained no exogenous zinc ion.
Both buffers were supplemented with 15 mM imidazole, a mixture of antiproteases and DNase.
Wild-type and mutant FICD/HYPE were expressed and purified as EfFIC, except that the lysis buffer was supplemented with 1 mM dithiothreitol (DTT) and 0.02 % Triton X-100 and the other buffers were supplemented with 1 mM DTT.
H-Ras purification buffers were supplemented with 2 mM MgCl2.
Wash buffers were supplemented with 0.1% SDS.
All buffers were supplemented with protease and phosphatase inhibitors (Sigma).
These were supplemented by mechanical pressure gauges.
Lysis and wash buffer were supplemented with 5 mM imidazole and elution buffer supplemented with 300 mM imidazole.
When optical tweezers were used, the ATP buffer was supplemented with NeutrAvidin-functionalized silica microspheres.
AP1 lysis buffer was supplemented with 2.5%PVP400.
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