Anti rabbit igg rhodamine

Double-label indirect immunofluorescence assay (IFA) was performed as previously described [34 (link)]. Briefly, cells grown on coverslips were fixed in 4% formaldehyde at 20 °C for 15 min and permeabilized in methanol at −20 °C for 10 min. The coverslips were incubated with an appropriate polyclonal antibody and a monoclonal antibody overnight at 4 °C and subsequently incubated with anti-rabbit IgG-rhodamine (Invitrogen Cat No. 31670, Waltham, MA, USA; 1:200 dilution) and anti-mouse IgG-FITC (Sigma-Aldrich Cat No. F0257-1ML; 1:100 dilution) at room temperature for 1 h. Slides were prepared with UltraCruz mounting medium (Santa Cruz Biotechnology) and visualized using an Eclipse fluorescence microscope (Nikon, Tokyo, Japan). Densitometric analysis of the immunofluorescence signal was performed using the ImageJ software (NIH, Bethesda, MD, USA).

Cells that were grown on coverslips under the indicated conditions were fixed in 4% formaldehyde at 20°C for 15 min and permeabilized in methanol at −20°C for 10 min. The coverslips were then incubated at 20°C for 3 h with anti-HCV core protein polyclonal (catalog no. ab58713; Abcam) (1:200) and anti-HBx monoclonal (catalog no. sc-57760; Santa Cruz) (1:500) antibodies (Fig. 1C and D; see also Fig. S1D at https://dv.pusan.ac.kr/SynapDocViewServer/viewer/doc.html?key=402882e382e741b101845c07289e5ac8&convType=html&convLocale=ko&contextPath=/SynapDocViewServer/) and with anti-HA polyclonal (catalog no. ab9110; Abcam) (1:200) and anti-HCV core protein monoclonal (catalog no. ab-2740; Abcam) (1:200) antibodies (Fig. 2D). The cells were then incubated with anti-rabbit IgG–rhodamine (catalog no. 31670; Invitrogen) (1:200 dilution) and anti-mouse IgG–fluorescein isothiocyanate (FITC) (catalog no. F0257-1ML; Sigma-Aldrich) (1:100 dilution) antibodies at 20°C for 1 h. The slides were prepared with UltraCruz mounting medium (catalog no. sc-24941; Santa Cruz Biotechnology) and visualized using an Eclipse fluorescence microscope (Nikon).