GSM3736834: SUM159_SMAD3_rep1; Homo sapiens; ChIP-Seq
After sonication, clarified nuclear lysates were pre-cleared with Protein A/G agarose for 1 hour. Pre-cleared lysates were then immunoprecipitated with 10 μg SMAD3 antibody (ab28379) overnight at 4°C. Protein A/G agarose blocked with BSA were added for 2 hours the next day, and beads were washed extensively. Protein/DNA complexes were eluted from the beads with 100 mM NaHCO3 and 1% SDS, and de-crosslinking was carried out overnight at 65°C. RNA and protein were digested with RNase A and Proteinase K, respectively. DNA was purified using phenol/chloroform/isoamyl alcohol (25:24:1) and precipitated overnight at -20°C. Libraries of ChIP DNA were prepared using the KAPA Hyper Prep Kit (KAPA Biosystems) according to manufacturer's instructions, and samples were multiplexed with Illumina adapters (NEB).
GEO Accession
GSM3736834