The mixture was incubated at 30 °C under continuous shaking.
All steps were conducted at 37°C under continuous shaking.
The reporter signal, streptavidine PE (Becton Dickinson, San Jose, CA, USA), was added and the bead mixture was incubated for 30 min under continuous shaking.
Mixes of beads were incubated for 2 hours with a standard, samples, or blank in a final volume of 50 µl for 2 hours under continuous shaking.
The reaction vials were subsequently kept at room temperature under continuous shaking (around 500 rpm).
The homogenate was then thoroughly mixed by vortexing and incubated for 10 min under continuous shaking.
The remaining 40 μl of Protein G Sepharose suspension were incubated with PBS as control in a glass vial for one hour at 4°C under continuous shaking.
Liver slices were incubated in William's E Medium (Lonza, Verviers, Belgium) in 6-well plates at 37°C/5% CO2/80% O2 under continuous shaking.
Incubation at 56°C with continuous shaking was continued for an hour.
Then, cells were washed and incubation was continued for 15 min at 37°C with continuous shaking.
A continuous shaking of the air and earth followed, then darkness, fog, and, later, out of the mushroom cloud, "a shower of most peculiar rain.
