Primary rabbit anti cd63

Standard immunoelectron staining with anti-CD63 antibody was performed as previously described [12 (link)]. The fixed exosomal preparations were placed on a carbon Formvar-coated 200-mesh nickel grid and incubated for 30 minutes. The grid was then quenched with 1 M ammonium chloride for 30 min and blocked with 0.4% BSA in PBS for 2 hrs. The grid was washed with PBS and incubated with primary rabbit anti-CD63 (1:100 Santa Cruz Biotechnology, Inc., Santa Cruz, CA) for 1 hr. The grid was then washed with ddH₂O and PBS, and 1.4 nm anti-rabbit nanogold (1:1000, Nanoprobes, Inc.) were dropped in blocking buffer for 1 hr. After enhancement with HQ Silver (gold enhancement reagent, Nanoprobes, Inc.), the samples were dried prior to observation in a transmission electron microscope (JEOL JEM 1230, Peabody, MA). TEM sample preparation and imaging were performed at the Electron Microscopy and Histology Core Laboratory at Augusta University (www.augusta.edu/mcg/cba/emhisto/).

Standard immunoelectron staining with anti-CD63 antibody was performed as
previously described (18 (link)). The fixed
exosomal preparations were placed on a carbon Formvar-coated 200-mesh nickel
grid and incubated for 30 minutes. The grid was then quenched with 1 M ammonium
chloride for 30 minutes and blocked with 0.4% BSA in PBS for 2 hours. The grid
was washed with PBS and then incubated with primary rabbit anti-CD63 (1:100
Santa Cruz Biotechnology, Inc., Santa Cruz, CA) for 1 hour. The grid was then
washed with ddH₂O and PBS, and drops of 1.4 nm anti-rabbit nanogold
(1:1000, Nanoprobes, Inc.) were applied in blocking buffer for 1 hour. After
enhancement with HQ Silver (gold enhancement reagent, Nanoprobes, Inc.), the
samples were wicked dry and allowed to air dry prior to observation in a
transmission electron microscope (JEOL JEM 1230, Peabody, MA). TEM sample
preparation and imaging were performed at the Electron Microscopy and Histology
Core Laboratory at Augusta University (www.augusta.edu/mcg/cba/emhisto/).