WB
Western blot analysis of lysates from HeLa cells using Phospho-Histone H3-S28 Rabbit mAb at 1:6000 dilution incubated at room temperature for 1.5 hours. HeLa cells were treated with Nocodazole(100 ng/mL) at 37℃ for 17 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 30 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Exposure time: 1 s.
WB
Western blot analysis of various lysates using Phospho-Histone H3-S28 Rabbit mAb at 1:6000 dilution incubated overnight at 4℃. NIH/3T3 and C6 cells were treated with Nocodazole (100 ng/mL) at37℃ for 17 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 30 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Exposure time: 20 s.
IP
Immunoprecipitation analysis of 600 μg extracts of HeLa cells treated with Calyculin A (100nM, 30min)using 5 μg Phospho-Histone H3-S28 Rabbit mAb. Western blot was performed from theimmunoprecipitate using Phospho-Histone H3-S28 Rabbit mAb at a dilition of 1:2000.