Aurein 3.3 from Ranoidea raniformis (Southern bell frog) (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P82396","term_id":"20137316","term_text":"P82396"}}P82396 AUR33_RANRN [https://www.uniprot.org/uniprot/{"type":"entrez-protein","attrs":{"text":"P82396","term_id":"20137316","term_text":"P82396"}}P82396]; sequence GLFDIVKKIAGHIVSSI) and uperin 3.5 from Uperoleia mjobergii (Mjoberg’s toadlet) (UniProt ID {"type":"entrez-protein","attrs":{"text":"P82042","term_id":"7994721","term_text":"P82042"}}P82042 | UPE35_UPEMJ [https://www.uniprot.org/uniprot/{"type":"entrez-protein","attrs":{"text":"P82042","term_id":"7994721","term_text":"P82042"}}P82042]; sequence GVGDLIRKAVSVIKNIV-NH2) were purchased from GL Biochem (Shanghai) Ltd. as lyophilized peptides, at >98% purity. The peptides were incubated in double distilled water (ddH2O) at a concentration of 5 mg/ml in 1.5-ml Eppendorf tubes at 20 °C. The optimal incubation time to produce ordered and dispersed fibers was monitored over time by negative-stain transmission electron microscopy of small aliquots, and determined to be 5 days. This time point was used for the preparation of cryo-samples.
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