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. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Nat Protoc. 2020 Apr 1;15(5):1690–1706. doi: 10.1038/s41596-020-0303-y

Figure 3. The experimental setup required for SoLVe-MS.

Figure 3

(i) Depending on the organism and complexity of the proteins embedded within the sample, as low as 50 μg of protein or as high as 10 mg of proteins embedded in membranes are required per experiment. The mechanism of sonication is unclear, however, before sonication bacterial inner membrane tubes are intact. Post-sonication the tips are removed suggesting sonication allows ingress of MS compatible buffer. Cryo-EM inset scale bar is 0.5 μm. (ii) Sonication is performed with the conditions stated. After sonication, and concentration of the vesicles, this preparation can be injected into an MS instrument. SoLVe-MS has been successfully attempted on a Thermo Q-Exactive UHMR and a Waters Synapt HDMS. Exact conditions are specified in the protocol. The overall suggestion is however to use high energy conditions. Cryo-EM inset scale bar is 0.5 μm. (iii). The entire process should take 2-3 days.