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![[Figure 2]](he5693fig2mag.jpg)
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Figure 2
Procedure for vitrification of Gab1–SHP2 complex crystals using the ThermoFisher Vitrobot Mark IV. (a, b, d) All-grid atlas views of crystal specimens (scale bar 450 µm) with enlarged views of corresponding single grid squares (insets, box edges 150 µm). (a) Gab1–SHP2 crystal sample (viscous slurry containing 29% PEG 3350) by double-sided blotting for 12 s at 4°C and 95% humidity. (b) Gab1–SHP2 sample upon increasing the double-sided blotting time to 24 s. (c) Schematic drawing demonstrating the optimized vitrification setup for preparing protein crystals for electron diffraction experiments. Complex crystals were harvested from the crystallization drops and applied directly to the front face of the EM grid. Crystallization buffer, diluted 1:1 with water, was applied to the back face of the grid. The plunger was mounted with a Parafilm disk on the side facing the crystals and with blotting paper on the side facing the buffer solution. (d) Gab1–SHP2 specimen prepared using the optimized setup, revealing microcrystals on the grid. (e) Low-magnification cryo-EM image of Gab1–SHP2 complex crystals observed after sample vitrification using the optimized setup. Two long rod-shaped crystals are visible on the grid. |
![[Figure 2]](he5693fig2.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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