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. Author manuscript; available in PMC: 2013 Jan 21.
Published in final edited form as: Methods Mol Biol. 2012;796:133–174. doi: 10.1007/978-1-61779-334-9_8

Figure 8. Hanging Drop Vapor Diffusion.

Figure 8

Step 1: Prepare the reservoir solution by mixing appropriate volumes of filtered solutions in a single well of a 24-well crystallization tray. The final volume of reservoir solution is typically 1 mL. Step 2: Pipette equal volumes of the purified protein and the reservoir solution onto a siliconized cover slip. There is no need to mix the two components. A common starting volume for the hanging drop is 2 mu;L (1 μL protein + 1 μL of reservoir solution), but can range from <1μL to 10 μL. Step 3: Carefully invert the coverslip without disturbing the droplet and place over the corresponding well. Gently press down on the coverslip to ensure an air-tight seal between the coverslip and the well.