EV71 Capsid Protein VP-1 Antibody Protocol

Mouse monoclonal antibody against EV71 capsid protein VP-1 was purchased from Abcam (Cambridge, UK). Rabbit monoclonal antibody to AP2M1 (Abcam), Rabbit polyclonal antibodies to FCHO2 (ProSci, Poway, CA, USA), eIF4G (Cell Signaling Technology, Danvers, MA, USA) and Enterovirus 71 2C and 3C (GeneTex, Irvine, CA, USA), Mouse monoclonal antibodies against clathrin (CLTC, Santa Cruz Biotechnology, Dallas, Texas, USA) and dynamin (Santa Cruz Biotechnology) were used in Western-blot detection of the respective proteins. The secondary antibody conjugated to Allophycocyanin (APC) was from BD Biosciences (Franklin Lakes, CA, USA). AlexaFluor594-Conjugated AffiniPure Goat Anti-Mouse IgG (H + L) was from ZSGB-BIO (Beijing, China). Fluoroshield Mounting Medium with DAPI was from Abcam. AlexaFluor 488 conjugated transferrin from human serum and AlexaFluor 594 conjugated cholera toxin subunit B were from ThermoFisher scientific (Waltham, MA, USA). Chloropromazin and dynasore were purchased from Selleckchem (Shanghai, China). Guanidine hydrochloride (GnHCl) was from MedChem Express (Monmouth Junction, NJ, USA). All drugs were prepared according to the manufacturer’s instructions under sterile conditions.