Rabbit antibody against ha

CoraLite^®488-conjugated CD9 monoclonal antibody (Catalog number: CL488-60232), HRP-conjugated GFP monoclonal antibody (Catalog number: HRP-66002), rabbit IgG control polyclonal antibody (Catalog number: 30000-0-AP), mouse antibody against β-actin (Catalog number: 66009-1-Ig), and rabbit antibodies against VPS25 (Catalog number: 15669-1-AP), GFP (Catalog number: 50430-2-AP), HRS (Catalog number: 10390-1-AP), and Tsg101 (Catalog number: 28283-1-AP) were purchased from ProteinTech Group (Wuhan, China). Mouse antibody against CD63 (Catalog number: sc-5275) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit antibodies against CD9 (Catalog number: 98327S) and calnexin (Catalog number: 2679) were purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit antibody against HA (Catalog number: H6908) and mouse antibody against FLAG (Catalog number: F3165) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit antibody against EV71 VP1 (Catalog number: GTX132339) was purchased from GeneTex, Inc. (Irvine, CA, USA). Rabbit antibody against EV71 3C (Catalog number: A10003) was purchased from ABclonal Technology (Wuhan, China). Mouse antibody against EV71 3A was kindly provided by Dr. Yongbo Yang of Central China Normal University.

Epimastigotes were washed with PBS (pH 7.4) and fixed with 4% paraformaldehyde and 0.05% glutaraldehyde in PBS (pH 7.4) for 1 h on ice. Samples were processed for cryo-immunoelectron microscopy at the Molecular Microbiology Imaging Facility, Washington University School of Medicine. HA-fused protein localization was detected with a rabbit antibody against HA (Sigma) at a 1:200 dilution for 1 h, at RT and goat anti-rabbit conjugated to 18 nm colloidal gold as a secondary antibody for 1h at RT.