Dylight488 labeled goat anti rabbit igg

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.

Antibodies were obtained from the following sources: GLUT-1 (07-1401), Claudin-5 (ABT45) were purchased from Millipore (Temecula, CA, United States); GAPDH (ab8245), CD-31 (ab24590), Dylight-488-labeled goat anti-rabbit IgG (Abacm,ab96899) and Dylight-594-labeled goat anti-mouse IgG (Abacm,ab96873) were purchased from Abcam (San Francisco, CA, United States); AQP-4 (A5971) and GFAP (G6171) from Sigma (St Louis, MO, United States); Occludin from Thermo Fisher (Waltham, MA, United States); ZO-1 (GTX108613) from GenTex (Irvine, CA, United States); and VE-cadherin (ARG20566) from Arigo (Hsinchu, Taiwan, China).