Goat anti rat igg h l alexa fluor 488 conjugates

Each mouse was i.p. injected with 200 μl of sterile BrdU solution (10 mg/ml). After 2 hours, livers were harvested to prepare single-cell suspensions in PBS using 40 μm cell strainers. Cells were slowly added to 70% ethanol at -20°C with continuous vortex followed by 30-minute incubation on ice. To denature DNAs, re-suspended cells were treated with 2N HCl containing 0.5% Triton X-100 and incubated at RT for 30 minutes. After washing with PBS containing 1% BSA and 0.05% Tween-20, cells were incubated with 1:100 diluted rat anti-BrdU antibodies (clone BU1/75, MCA2060, AbD Serotec) at RT for 45 minutes followed by incubation with 1:200 diluted goat anti-Rat IgG (H+L) Alexa Fluor^®488 conjugates (A-11006, Life Technologies). After washing, cells were stained with PBS containing 10 μg/ml of PI before flow cytometry analysis.