Lysate-to-grid: Lessons learned from cryo-EM of endogenous protein complexes

The success of any structural biology project is dependent on achieving sufficient purity and concentration of the sample-of-interest. Samples for cryo-EM generally have lower requirements for purity and concentration compared to other structural methods, making it a highly attractive approach for determining high-resolution structures of particles isolated directly from their native sources. Here, I will describe i) our general approach to preparing cryo-EM specimens of native purifications, ii) successes and failures in our efforts, and iii) ongoing efforts to teach cryo-EM newcomers about calibrating expectations for their own projects.