Anti rabbit or anti mouse igg antibodies

Example 13

Murine liver tissues were dissected and then cryoprotected in 30% sucrose for the preparation of paraffin-embedded samples. Paraffin samples for Hematoxylin and Eosin (H&E) staining were prepared. After removing paraffin, rehydrated samples were incubated with monoclonal antibodies to human hepatocytes (Hep par 1) and HCV NS5A (Santa Cruz, Calif., USA), which were diluted at a ratio of 1:50, at 4° C. overnight. After washing 3 times, the samples were incubated with anti-rabbit or anti-mouse IgG antibodies (Invitrogen, Carlsbad, Calif., USA), which were conjugated with Alexa Fluor 488 or Alexa Fluor 546, which were diluted at a ratio of 1:100, for 2 hours. The nuclei were stained with DAPI for 5 minutes and analyzed using a confocal microscope.

Example 23

HCV replicon Cells were seeded onto the coverslips within a 12-well plate to about 70% confluence for 24 hours. For fixation, the cells were washed 3 times with PBS and fixed with 4% paraformaldehyde solution for 20 minutes at room temperature. Then, the cells were washed again 3 times with PBS. For providing permeability, the cells were incubated in PBS containing 0.5% Triton X-100 for 15 minutes, and washed 3 times with PBS before incubating with primary antibody. The primary antibody was diluted to a desired concentration with 1×PBS containing 3% BSA, in general at a ratio of 1:100, to prevent non-specific binding of the antibody. After overnight incubation, the cells washed 3 times with PBS, and incubated with secondary antibody, which was conjugated with anti-rabbit or anti-mouse IgG antibodies (Invitrogen, Calif., USA), which was conjugated with Alexa 488 or Alexa 546 diluted with PBS containing 3% BSA at a ratio of 1:250, for 2 hours. The cells were counterstained with DAPI and mounted onto glass slides (Vector Mount, Calif., USA). The thus-prepared samples were photographed using the Zeiss confocal microscope (Zeiss, Germany).