1a,e–g and 2e,f were acquired on verso Refeyn OneMP mass photometer with per 10.8 ? 2.9 ?m 2 (128 https://www.datingranking.net/it/tinychat-review/? 35 pixels) field of view. The microscope used onesto acquire the momento sopra Figs. 1a,e–g and 2e,f was custom-built with a 9.4 ? 6.2 ?m 2 field of view (138 ? 88 pixels with per binned pixel size of 70.3 nm) and is similar sicuro that described previously 18 . The custom-built setup is illustrated con Supplementary Fig. 15 and the differences onesto the setup used mediante our previous sistema are highlighted durante the figure caption.
Dynamic mass photometry
Sicuro acquire dynamic mass photometry movies, the SLBs were placed on the sample tirocinio onesto optimize the focus of the microscope. After locking the focus of the microscope, WT or ?PRD was added onesto the SLB by replacing 3–6 ?l buffer from the silicone gasket with 3–6 ?l (total libro, 60 ?l) 200 nM WT or ?PRD (mediante reaction buffer) solution and mixing well with verso micropipette to achieve a final concentration of 10–20 nM. Datazione acquisition was started ?10 s after the additif of protein. Images were collected at 994 Hz and saved after the binning of pixels into blocks of 4 ? 4 and the binning of frames into groups of 3, resulting in an effective frame rate of 331 Hz and verso final pixel size of 84.4 nm (for the giorno used sopra Extended Data Figs. 2, 3, 8 and Supplementary Figs. 3 and 13 the pixel size was 77.4 nm).
5–10 s) once every minute (20,000 frames) to readjust the microscope focus esatto account for drift over time before resuming acquisition. This resulted per sets of multiple 1 min movies for each SLB and sample combination. For experiments on the effect of GTP, 3–4 movies of WT (10–20 nM) were recorded as described above, after which image acquisition was briefly paused and 1.2 ?l GTP (50 mM) was added (total gasket testo, 60 ?l), followed by missaggio esatto obtain verso final concentration of
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