Ab28629

Nuclear lysates were prepared as previously described^{31 (link)}. Nuclear extracts were separated on either 8%, 10% or 20% SDS-PAGE gels. Blots were probed with primary antibodies (H2A-Ub[K119] [Cell Signaling Technology], H2A [Cell Signaling Technology], H2B-ub [EMD Millipore], H2B [Cell Signaling Technology #12364, clone D2H6], H3 [Cell Signaling Technology], H4 [Cell Signaling Technology], HA [Cell Signaling Technology #2367, clone 6E2], TRIM37 [Abcam ab95997, or custom made by 21^st Century Biochemicals against a synthetic peptide corresponding to amino acids 444–460 of the human protein followed by affinity purification] or], RNF2 [Abcam ab28629], and α-tubulin [in-house]) overnight at 4°C, washed five times in TBS plus 0.1% Tween (TBST) and then incubated with the appropriate HRP-conjugated secondary antibody for 1 h at room temperature. Membranes were washed five times in TBST and visualized on autoradiography film after incubating with ECL reagent (Supersignal West Pico or Supersignal West Femto; Thermo Scientific). Immunoblots were quantified using Image J software version 1.47v (NIH).