Alexa488 labeled goat anti mouse igg

Cells were grown on coverslips, washed with PBS and fixed for 20 min in 4% paraformaldehyde (PFA). Cells were then permeabilized in 0.5% Triton X-100 for 3 min. After blocking, cells were immunostained for 1 hr with a primary antibody, and after subsequent washes the cells were incubated for 1 hr with secondary fluorescent antibodies. Primary antibodies: anti-SRSF2 (Sigma). Secondary antibodies: Alexa488-labeled goat anti-mouse IgG (Abcam) and Alexa594-labeled goat anti-mouse. Nuclei were counterstained with Hoechst 33342 and coverslips were mounted in mounting medium.

Cells were grown on 18 mm coverslips, washed with phosphate-buffered saline (PBS) and fixed for 20 min in 4% PFA. U2OS Tet-On cells were then permeabilized in 0.5% Triton X-100 for 2 min, after which they were blocked in bovine serum albumin (BSA). HFF-1 and HepG2 were permeabilized twice in 0.1% Triton X-100 for 5 min. Cells were immunostained for 1 hr with a primary antibody, and after subsequent washes the cells were incubated for 45 min with secondary fluorescent antibodies. Primary antibodies: Mouse anti-Serine and Arginine Rich Splicing Factor 2 (SRSF2) (which marks SRRM2 [52 (link)]), rabbit anti-SON (Sigma), rabbit anti-Heterogeneous Nuclear Ribonucleoprotein K (hnRNPK) (Abcam), rabbit anti-SRRM2 (Abcam) and rabbit anti-Serine and Arginine Rich Splicing Factor 7 (SRSF7) (Santa Cruz). Secondary antibodies: Alexa647-labeled goat anti-mouse IgG, Alexa594-labeled goat anti-mouse IgG, Alexa647-labeled donkey anti-rabbit IgG (Life Technologies, Thermo Fisher Scientific), dyLight488-labeled goat anti-rabbit IgG and Alexa488-labeled goat anti-mouse IgG (Abcam). The cells were mounted in mounting medium.

Cells were grown on coverslips, washed with PBS and fixed for 20 min in 4% PFA. Cells were then permeabilized in 0.5% Triton X-100 for 2.5 min. Cells were washed twice with PBS and blocked with 5% BSA for 20 min and immunostained for 1 h with a primary antibody. After three washes with PBS, the cells were incubated for 1 h with secondary fluorescent antibodies. Primary antibodies: mouse anti-G3BP1 (Abcam) and mouse anti-Hedls (Santa Cruz). Secondary antibodies: Alexa488-labeled goat anti-mouse IgG (Abcam). Nuclei were counterstained with Hoechst 33342 (Sigma) and coverslips were mounted in mounting medium.