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The motility, curvilinear velocity (VCL) and straightness (STR) values did not decrease significantly during 60 min of equilibration neither in equilibrated nor thawed groups.
After 10 min of equilibration, they were removed for volumetric analysis.
After 10 min of equilibration, 0.5 g/L of enzyme was added to the solution.
It was found that the absorbance value was found to be constant after 20 min of equilibration.
The second stage of equilibration with variamine blue in presence of sodium acetate was found to be completed in about 10 min of equilibration.
After 10 min of equilibration at 37°C, bacteria were treated with dye alone or with increasing concentrations of squalamine; CTAB at 58 µg/mL (i.e. 160 µM)[40] being used as positive control of membrane permeabilisation.
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After the 30 min period of equilibration, the contractile response to depolarizing potassium solution was assessed as a test for viability.
After a 10 min period of equilibration in start buffer (0.05% v/v TFA/water), peptides were eluted using a gradient formed from start buffer to 0.05/19.95/80.00 (v/v/v) TFA/water/acetonitrile in 240 min at a flow rate of 1 mL/min.
After isoelectric focusing the IPG strips were equilibrated for 15 min in 10 mL of equilibration buffer (6 M urea, 30% [v/v] glycerol, 2% [w/v] SDS (sodium dodecyl sulfate), 75 mM Tris, 0.002% [w/v] bromophenol blue) containing 1% (w/v) DTT and subsequently for 15 min in 10 mL of equilibration buffer containing 2.5% (w/v) iodoacetamide.
Subsequently strips were equilibrated for 10 min in 10 ml of equilibration buffer (6 M urea, 30% [vol/vol] glycerol, 2% [wt/vol] SDS (sodium dodecyl sulfate), 75 mM Tris, 0.002% [wt/vol] bromophenol blue) containing 1% (wt/vol) DTT and subsequently for 10 min in 10 ml of equilibration buffer containing 2.5% (wt/vol) iodoacetamide.
The strips were equilibrated for 15 min in 10 ml of equilibration solution (0.375 M Tris HCl, pH 8.8, 6 M urea, 20% [v/v] glycerol and 2% [w/v] SDS), with 2% (w/v) DTT and for 15 min in 10 ml of equilibration solution containing 2% (w/v) iodoacetamide.
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