Goat anti mouse igg h l alexa fluor 488 preabsorbed

Doxycycline-induced TSA^shNS and TSA^shTrex1 cells were exposed to 0 Gy, 8 Gy or 20 Gy. 24 h after radiation, cells were fixed for 10 min at room temperature in 4% PFA in PBS. Cells were permeabilized with 0.1% Tween 20 and 0.01% Triton X-100 for 20 min, followed by blocking with 1% BSA, 22.52 mg ml⁻¹ glycine in PBST (0.1% Tween 20 in PBS) for 30 min at RT. Cells were incubated with a primary anti-dsDNA antibody (Abcam. Cat # ab27156; 1:1,000) overnight at 4 °C. After three washes, samples were incubated with a secondary antibody (goat anti-mouse IgG H&L—Alexa fluor 488 preabsorbed, Abcam, Cat # ab150117; 1:200) for 1 h at RT. After washes, mounting medium containing DAPI was used (Vector Laboratories, Cat # H-1500) to counterstain nuclei. Images were obtained using an EVOS FL microscope from Life Technologies.