Alexa fluor 555 conjugated anti rabbit igg antibody

RCC cells were plated in 12-well plates at 30% confluence and allowed to grow for 24 h. Then, the cells were fixed with 10% paraformaldehyde solution for 15 min at room temperature, permeabilized with 0.4% Triton X-100 in PBS for 5 min, and then blocked with 1% BSA in PBS for 1 h at 37 °C. The blocked cells were incubated with anti-FLAG antibody (1:100, Merck Millipore) and anti-YAP antibody (1:100, Cell Signaling Technology) overnight at 4 °C, followed by incubation with Alexa Fluor 488-conjugated anti-mouse IgG antibody and Alexa Fluor 555-conjugated anti-rabbit IgG antibody (1:100, Invitrogen, Carlsbad, CA) for 2 h. Nuclear staining of cells was conducted using 4,6-diamidino-2-phenylindole (DAPI). Representative images were acquired using an fluorescence microscope (IX70, Olympus, Japan).

RCC cells were plated in laser confocal special culture dishs at 30% confluence and treated with indicated reagents at indicated concentration for 48 h. Then, the cells were fixed with 4% paraformaldehyde solution for 15 min at room temperature, permeabilised with 0.4% Triton X-100 in PBS for 5 min, and then blocked with 1% BSA in PBS for 1 h at 37 °C. The blocked cells were incubated with anti-FLAG antibody (1:100, Sigma) and anti-FAK antibody (1:100, abcam) overnight at 4 °C, followed by incubation with Alexa Fluor 488-conjugated anti-mouse IgG antibody
and Alexa Fluor 555-conjugated anti-rabbit IgG antibody (1:100, Invitrogen, Carlsbad, CA) for 2 h. Nuclear staining of cells was conducted using 4,6-diamidino-2-phenylindole (DAPI). Representative images were acquired using the Leica Microsystem.