Anti ezh2 rabbit polyclonal antibody

Quantitative ChIP assays were performed as described previously [47 (link), 48 (link)]. Cross-linked chromatin prepared from the indicated cells was precipitated by normal rabbit IgG (Wako 148–09551), an anti-EZH2 rabbit polyclonal antibody (Millipore 07–689), or anti-trimethyl-Histone H3K27 rabbit polyclonal antibody (Millipore 07–449) with Protein G Sepharose 4 Fast Flow (GE Healthcare, Chicago, IL, USA). The primers used in the present study are indicated in Supplementary Table S4. Each series of experiments was conducted at least three times.

Cells were lysed in EBC buffer [9 (link)] or CHAPS buffer (10 mM CHAPS, 50 mM pH 7.5 Tris-HCL, 100 mM NaCl, 10 mM MgCl₂, and 1 mM EDTA) for the immunoblotting of NTRK1, and protease/phosphatase inhibitors were added to the buffers [9 (link)]. Western blotting experiments were performed as previously reported [45 (link)]. After being transferred to an Immobilon-P membrane (Millipore, Billerica, MA, USA), proteins were reacted with anti-EZH2 rabbit polyclonal antibody (Millipore 07–689), anti-BMI1 mouse monoclonal antibody (Millipore 229F6), anti-RING1B antibody [46 (link)], anti-trimethyl-Histone H3 rabbit polyclonal antibody (Millipore 07–449), histone H3 rabbit polyclonal antibody (Abcam ab1791, Cambridge, United Kingdom), anti-TrkA (NTRK1) rabbit monoclonal antibody (Cell Signaling Technology 14G6, Beverly, MA, USA), or anti-β-actin antibody (Sigma-Aldrich).