Anti ezh2 antibody

Cardiomyocytes were lysed in RIP lysis buffer, following incubation with RIP buffer containing magnetic beads conjugated with anti-EZH2 antibody (Millipore, USA) or negative control IgG. Anti-SNRNP70 (Millipore, USA) was used as positive control for the RIP procedure. The samples were incubated with Proteinase K with shaking to digest the protein and then immunoprecipitated RNA was isolated. The RNA concentration was measured using a NanoDrop (Thermo Scientific) and the RNA quality assessed using a bioanalyser. Furthermore, purified RNA was subjected to real-time PCR to determine the presence of binding targets using respective primers.

Cardiomyocytes were lysed in RIP lysis buffer, following incubation with RIP buffer consisting of 150 mM KCl, 25 mM Tris pH 7.4, 5 mM EDTA, 0.5 mM DTT, 0.5% NP40, 100 U/ml RNAase inhibitor SUPERASin. The RIP buffer contained magnetic beads conjugated with anti-EZH2 antibody (Millipore, USA) or negative control IgG. The samples were incubated with Proteinase K and then immunoprecipitated RNA was isolated. Purified RNA was subjected to real-time PCR to determine the presence of binding targets using respective primers.

RNA immunoprecipitation (RIP) assays were performed with Magna RIP™ RNA‐Binding Protein Immunoprecipitation Kit (Millipore) according to the manufacturer's instructions. We obtained an anti‐EZH2 antibody for RIP tests from Millipore. After incubating RNase‐treated RIP lysates for 1 hour  at 37 °C, we detected co‐precipitated RNAs using RT‐PCR.