Western Blotting Protocol with Antibodies

Western blotting was performed using standard methods as described previously (Albisetti et al., 2017 (link)), with the exception that proteins were transferred to nitrocellulose membranes (Genesee Scientific, Prometheus #84-875). Primary antibodies were used at the following dilutions: rabbit anti-KIN-E 1:2000; rabbit anti-GST 1:1000 (Welch lab); mouse anti-β-tubulin E7 1:10,000 (Developmental Studies Hybridoma Bank, University of Iowa). Secondary antibodies used were goat anti-rabbit AF790 (ThermoFisher Scientific A11367) and goat anti-mouse AF680 (ThermoFisher Scientific A21058), both diluted at 1:10,000. Images were taken using the Odyssey imaging system (Li-Cor Biosciences).