Murine anti flag antibody

FLAG® Immunoprecipitation assay eluate samples were detected using murine anti-FLAG antibody (1:4000) (Sigma-Aldrich) or anti-Chloramphenicol Acetyl Transferase (rabbit IgG fraction) (1:6000) (Sigma-Aldrich). The secondary antibodies employed were goat anti-mouse HRP (1:10 000) (Sigma-Aldrich) and anti-rabbit-HRP (1:10000) (Sigma-Aldrich), respectively. The loading control, β-actin, was detected employing the rabbit anti-β-actin-HRP antibody (1:10000) (Sigma-Aldrich). Experiments were performed in triplicate, three times.

Anti-CSDE1 rabbit antibody was obtained from Bethyl Laboratories (catalog no. A303-159A). ACBD3-specific rabbit antibody (catalog no. HPA015594), murine anti-FLAG antibody (catalog no. F3165), and beta actin murine monoclonal antibody (catalog no. A1978) were purchased from Sigma-Aldrich. Murine monoclonal anti-VP1 antibody was obtained from Leica (NCL-ENTERO). Anti-RACK1 mouse antibody was purchased from BD Biosciences (catalog no. 610177). Rabbit polyclonal anti-3C antibody was generated by immunization with recombinant 3C protein. Secondary antibodies conjugated to horseradish peroxidase were purchased from Thermo Fisher Scientific (catalog no. 31466, 31430). Alexa Fluor 488-conjugated goat anti-rabbit antibody was obtained from Life Technologies (catalog no. A11008). Chemical inhibitor Dynasore was purchased from Merck Millipore, and C75 and itraconazole were purchased from Sigma-Aldrich.