AIBP was produced in a baculovirus/insect cell system to ensure posttranslational modification and endotoxin-free preparation as described in Choi et al. (2018) (link) and Woller et al. (2018) (link). Bulk production of AIBP was ordered from Selvita, Inc. wtAIBP and mutAIBP, mouse WT AIBP, and zebrafish WT AIBP (Fang et al., 2013 (link)) were cloned into a pAcHLT-C vector behind the polyhedrin promoter. The vector contains an N-terminal His-tag to enable purification and detection. Insect Sf9 cells were transfected with BestBac baculovirus DNA (Expression Systems) and the AIBP vector. After 4–5 d, the supernatant was collected to afford a baculovirus stock. Fresh Sf9 cells were infected with the AIBP-producing baculovirus; cell pellets were collected after 3 d, lysed, sonicated, and cleared by centrifugation, and the supernatants were loaded onto a Ni-NTA agarose column eluted with imidazole. Protein was dialyzed against saline, and concentration was measured. Aliquots were stored at −80°C.