Goat anti mouse igg dylight 800 conjugate

For analysis of whole cell lysates, samples were taken after subculturing for 4 h at 27°C (analysis of Inv expression) or 37°C (analysis of YadA expression). Pellets were resuspended in H₂O and Laemmli buffer (Bio-Rad) to obtain 5×10⁶ bacteria/ml. Samples were boiled for 10 min at 95°C before loading on the gel. After SDS-PAGE proteins were transferred onto nitrocellulose membranes. The membranes were blocked overnight with 5% milk powder in PBS at 4°C. Membranes were probed with antisera against Inv (rabbit anti-Inv, 1:2000), YadA (rabbit anti-YadA, 1:1000), OmpA (rabbit anti-OmpA, 1:1000), OmpF (rabbit anti-OmpF, 1:1000), BamB (rabbit anti-BamB, 1:1000) and RNA-Polymerase β subunit (mouse anti-RNA-Polymerase, Neoclone, 1:1000) and secondary antibodies goat anti-rabbit IgG DyLight 680 conjugate or goat anti-mouse IgG DyLight 800 conjugate (Thermo Fisher scientific). For detection of bands, the Odyssey imaging system (Li-Cor Biosciences) was used.