Immunocytochemistry analysis of RNF31 in HeLa cells. Samples were treated with PFA, permeabilized in 0.1% Triton X-100, blocked in 10% serum (45 min at 25°C), and incubated with polyclonal antibody (Product # PA5-115849) at a dilution of 1:200 (1 hr at 37°C). Secondary staining was applied with mouse anti-beta tubulin; AlexaFluor 594 conjugated goat anti-rabbit IgG (Red); AlexaFluor 88 conjugated goat anti-mouse IgG (Green) and DAPI (blue) using a dilution of 1:200 (1 hr at 37°C).