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Figure 4
Procedure for reducing perdeuterated MnSOD crystals by a full soak within a capillary. Arrows indicate the position of the crystal, while black lines indicate the positions of reservoir slugs supplemented with dithionite. (a) An MnSOD crystal flanked by dithionite-containing slugs within a capillary heat-sealed using wax. (b) Both ends of the capillary are opened by peeling off the wax (without heat). (c, d) One of the slugs is removed using a pipet attached to a gel-loading pipet tip. (e, f) Using a pipet, negative pressure is applied to engulf the crystal in the slug solution. (f) shows a crystal transitioning between redox states ∼20 s after initiation of the soak, while (g) depicts the full transition after 1 min. (h) The soaking solution is subsequently pipetted off or dried using wicks. During this process, it should be ensured that a slug is present within the capillary to ensure crystal hydration. (i) Another slug is added and the capillary is resealed. |
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