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![[Figure 4]](ic5104fig4mag.jpg)
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Figure 4
Practical applications of continuous carbon supports. (a)–(c) Avoiding aggregation by immobilizing the virus prior to the addition of a binding protein, (d)–(f) grid soaking with low-affinity receptor molecules. (a) An example of a virus-only sample distributed evenly across a holey grid. (b) Aggregates are observed on a grid after virus and non-antibody binding protein are mixed in solution. (c) Virus and non-antibody binding protein complexes are distributed evenly across a grid. Virus sample was applied to a lacey grid with a 3 nm continuous carbon support (Agar Scientific) and the excess solution was blotted away; the binding protein was then applied and the excess was washed away with buffer before blotting and plunge freezing. (d) Virus-only sample distributed evenly across a lacey grid with a 3 nm continuous carbon support (Agar Scientific). Owing to the low concentration of the sample and its intractability to concentration, multiple aliquots of virus sample were applied to the grid prior to blotting and plunge freezing. (e) Virus and 20 mM solution of receptor fragment. Excess virus sample was applied to a grid and blotted away before the concentrated receptor solution was applied. This was left to dwell for 30 s prior to blotting and plunge freezing. (f) EM density (2.8σ) and fitted model for a terminal sialic acid present in the low-affinity receptor fragment. Scale bars: (a)–(c) 100 nm; (d), (e) 50 nm. |
![[Figure 4]](ic5104fig4.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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