WB
Western blot analysis of various lysates using Phospho-Histone H3-T3 pAb at 1:1000 dilution. C6and HeLa cells were treated with Nocodazole (50 ng/mL) at 37℃ for 20 hours. C6 and NIH/3T3 cells weretreated with CIP(20uL/400ul) at 37℃ for 1 hour.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Exposure time: 1s.
WB
Western blot analysis of various lysates using Phospho-Histone H3-T3 pAb (AP0846) at 1:1000 dilution orHistone H3 antibody. Both 293T cells and Raw264.7 cells were treated with Nocodazole (100ng/mL) at 37℃ for 16 hours.C6 cells were treated with Nocodazole (50 ng/mL) at 37℃ for 20 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit.Exposure time: 1s.