Mcherry rabbit pab

Cell lysates were prepared from either mock- or virus-infected (MOI, 0.01) Vero E6 cells (1.2 × 10⁶ cells/well, 6-well format) after 24 hpi using passive lysis buffer (Promega) based on the manufacturer’s instructions. After centrifugation (12,000 × g) at 4°C for 30 min, proteins were separated with 12% SDS-PAGE and transferred to nitrocellulose membranes. Membranes were blocked for 1 h with 5% dried skim milk in 0.1% Tween 20 PBS (T-PBS) and incubated at 4°C overnight with the following specific primary MAbs or polyclonal antibodies (PAbs): N (mouse MAb 1C7C7), mCherry (rabbit Pab; Raybiotech), and Nluc (rabbit Pab, Promega). Then, membranes were incubated at 37°C for 1 h with goat anti-mouse IgG StarBright Blue 520 or anti-rabbit IgG Starbright Blue 700 (Bio-Rad) secondary antibodies. Tubulin was used as a loading control using an anti-tubulin hFAB rhodamine antibody (Bio-Rad). Proteins were detected using a ChemiDoc MP imaging system (Bio-Rad).

Cell lysates were prepared from either mock- or virus-infected (MOI, 0.01) Vero E6 cells (1.2 × 10⁶ cells/well, 6-well format) after 24 hpi using passive lysis buffer (Promega) based on the manufacturer’s instructions. After centrifugation (12,000 × g) at 4°C for 30 min, proteins were separated with 12% SDS-PAGE and transferred to nitrocellulose membranes. Membranes were blocked for 1 h with 5% dried skim milk in 0.1% Tween 20 PBS (T-PBS) and incubated at 4°C overnight with the following specific primary MAbs or polyclonal antibodies (PAbs): N (mouse MAb 1C7C7), mCherry (rabbit Pab; Raybiotech), and Nluc (rabbit Pab, Promega). Then, membranes were incubated at 37°C for 1 h with goat anti-mouse IgG StarBright Blue 520 or anti-rabbit IgG Starbright Blue 700 (Bio-Rad) secondary antibodies. Tubulin was used as a loading control using an anti-tubulin hFAB rhodamine antibody (Bio-Rad). Proteins were detected using a ChemiDoc MP imaging system (Bio-Rad).