Western blot analysis of extracts of 293T cells, using Phospho-POLR2A-S2 antibody (AP0749) at 1:1000 dilution. 293T cell lysate were treated by CIP (20ul CIP for each 400ul cell lysate) at 37℃ for 1 hour or treated by Doxorubicin (0.5uM) for 24 hours.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% BSA.|Detection: ECL Basic Kit (RM00020).|Exposure time: 1s.

Immunoprecipitation analysis of 200ug extracts of MCF-7 cells, using 3 ug Phospho-POLR2A-S2 pAb (AP0749). Western blot was performed from the immunoprecipitate using Phospho-POLR2A-S2 pAb (AP0749) at a dilition of 1:1000. MCF7 cells were treated by nocodazole (50 ng/mL) at 37℃ for 20 hours.

Immunofluorescence analysis of HeLa cells using Phospho-POLR2A-S2 antibody (AP0749) at dilution of 1:100. HeLa cells were treated by nocodazole (50 ng/mL) at 37℃ for 20 hours (left). Blue: DAPI for nuclear staining.

Western blot analysis of extracts of 293T cells, using Phospho-POLR2A-S2 pAb (AP0749) at 1:1000 dilution or POLR2A antibody (A2107).293T cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% BSA.|Detection: ECL Basic Kit (RM00020).|Exposure time: 1s.