Gold conjugated goat anti mouse igg

The purified Hcp1-loaded MVs were added to 230-mesh formvar/carbon-coated copper grids (Zhongjingkeji Tech., China), and negatively stained with 2% (m/v) uranylacetate for 30 sec. For immunogold labeling, Hcp1-loaded MVs was 1:20 diluted in NTE buffer (10 mM Tris-Cl, pH 7.0, 100 mM NaCl) and adsorbed onto 230-mesh formvar/carbon-coated nickel grids. After 5 min wash with TBS buffer (50 mM Tris-HCl, 150 mM NaCl, pH 7.5), the specimen was blocked with 3% (m/v) bovine serum albumin (BSA) in TBS for 45 min. Mouse-anti-Hcp1 antibody was 1:500 diluted in 1% (m/v) BSA/TBS and applied to the nickel grids for 1.0 h at room temperature. After washed three times with TBS, gold-conjugated goat-anti-mouse IgG (Sigma, USA) was added and incubated for 1.0 h at room temperature. Next, the grids were washed twice with TBS, once with water and followed by negatively stained with 2% (m/v) uranylacetate for 15 sec. Electron micrographs were recorded with a JEM1011 microscope (JEOL, Japan) at 100 kV acceleration voltage.