Figure 2
From: Tracking hantavirus nucleocapsid protein using intracellular antibodies
His-tag pull-down assay between intracellular scFvs and hantavirus N protein. Stably transfected Vero E6 cells, indicated as L13F3-scFv-ER and H34-scFv-ER, were infected with hantavirus 76-118 and L99 respectively. About 5 × 107 infected cells were collected and cell extracts incubated with Ni-NTA His-bind resin. After washing, bound proteins were eluted and concentrated by adding two volumes of ice-cold 100% ethanol and centrifuged at 10,000 × g for 10 min. Pellets were resuspended in 10 μl distilled water and analyzed by SDS-PAGE (A), Intracellular scFvs were analysized by immunoblotting with anti-penta-HIS (Qiagen) (B), and N proteins were blotted by N protein-specific mAbs L13F3 (C).