Western blot analysis: Cell lysates of MCF7 (20 ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human UBE2L6 (1:2000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

Paraffin embedded sections of human breast lobule tissue were incubated with anti-human UBE2L6 (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB)

Immunohistochemistry: Paraffin embedded sections of human breast lobule tissue were incubated with anti-human UBE2L6 (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB).