GSM4453187
GSM4453187: GFP_H3K27me3; Homo sapiens; ChIP-Seq
Chromatin immunoprecipitation was performed using the protocol described by (33); with minor modifications. Sonication of the samples was accomplished using a Bioruptor sonicator. Sheared-chromatin was diluted 10 fold in ChIP dilution buffer, supplemented with protease inhibitor. 50 μl of sheared chromatin was removed and stored at 4°C as control aliquot (INPUT).  Each sample was incubated overnight with antibodies (20 ug/ChIP) of interest. The following primary antibodies were used: H3K27me3 (07-449, Millipore), H3K4me3 (AB8580, Abcam), H3K36me3 (Ab9050, Abcam), H3K4me2 (AB7766, Abcam), H3K4me1 (AB8895, Abcam) and H3K27ac (AB4729, Abcam). Chromatin–Antibody complexes were then precipitated, washed eluted. DNA isolation was performed by phenol:chloroform:isoamyl alcohol, precipitated with NaCl, supplemented with glycogen, washed with EtOH and then treated with RNase I. Finally, DNA was purified with MinElute Kit. Libraries were prepared according to NEBNext UltraII DNA Library preparation kit (Illumina).
GEO
GSM4453187
GEO Accession
GSM4453187