Positive hits were scaled up in 24-well Linbro dishes by using the hanging-drop method of crystallisation, with crystallisation drops containing protein solution (1 μL) and precipitant reservoir (1 μL).
Crystals of the R517K protein were formed by using the hanging-drop method, by mixing 2 μl of the protein solution containing DNA with the ddTTP (10 mM) with 2 μl of reservoir solution containing 0.1 M KCl, 10 mM MgCl2, 50 mM Tris HCl, pH 8.5, and 30% PEG 400.
Crystals of the R517A mutant were formed by using the hanging-drop method, by mixing 2 μl of the protein solution containing DNA with 2 μl of reservoir solution containing 0.2 M ammonium acetate, 100 mM Hepes, pH 7, and 10% PEG 4000.
The apo-T151A protein (10 mg ml−1, 20 mM Tris pH 8.0) was crystallized by vapour diffusion using the hanging drop technique, by mixing the protein in a two-to-one ratio with reservoir solution at 20 °C.
Because this finding could reflect CTX leading to increased EB aggregation and, thus, larger EBs, we explored this possibility by growing individual EBs using the hanging drop method [24].
Embryoid bodies were generated using the hanging drop method, by aggregating 400 cells in 20 μl drops as described previously (Dang et al., 2002).
Remarkable improvement of the crystal quality was achieved by setting up crystallization conditions, at first established using the hanging drop vapor diffusion method, in a micro-batch experiment.
We generated embryoid bodies using the hanging drop method, and measured differentiation by analyzing the extent of the differentiated outgrowth upon plating the EBs.
Crystallization was performed using the hanging drop vapor diffusion method at 16°C by mixing equal volumes (1.0 μl) of protein solution (20 mg/ml) and reservoir solution.
Crystallization studies were completed using the hanging drop technique.
Human synovial MSCs were aggregated using the hanging drop technique.
