Goat anti rabbit horseradish peroxidase hrp antibody

To determine whether the E6_F47R protein was expressed correctly, CEFs and Vero and MRC-5 cells were infected with 10 PFU/cell FPE6_F47R and examined by Western blotting, as described previously [46 (link)]. The blotted nitrocellulose membranes were incubated overnight at 4°C with 1:100 dilution of the primary rabbit AbE6/Mu polyclonal antibody (M. Mueller, German Cancer Research Center, Heidelberg, Germany). After a 1-h incubation with a goat anti-rabbit horseradish peroxidase (HRP) antibody (1:2000 dilution; DakoCytomation, Carpinteria, CA, USA) and 2-h washes, the proteins were revealed using the ECL system (GE Healthcare, Buckinghamshire, UK). Cells infected with FPwt were used as the negative control, and the E6 protein as the positive control.

To assess the replication capacity of SARS-CoV-2 in human cardiomyocytes, a micro-plaque assay was performed. In this assay, Vero cells were inoculated with a serial dilution of the 42 or 72 h post-infection supernatant and incubated for viral adsorption at 37 °C for 2 h. The cells were then treated with MEM media containing 5% fetal bovine serum (FBS) and 1% agarose, followed by further incubation at 37 °C with 5% CO₂. After 24 h post infection, the cells were stained with a rabbit anti-nucleoprotein (NP) antibody and a goat anti-rabbit horseradish peroxidase (HRP) antibody (Dako). The SureBlue substrate (Merck) was added and allowed to develop for 10 min. The positive colonies formed were counted as plaque-forming units per milliliter (PFUs/mL), a measure of viral output.