GSM2574046: IGG-L11; Mus musculus; ChIP-Seq
ChIP was performed as previously described (Dere et al., 2001. BMC Genomics 12, 365; Lo and Matthews, 2012. Toxicol Sci 130(2), 349-61). Briefly, mouse livers (~100 mg) were homogenized in 1% formaldehyde and incubated at room temperature for 10 minutes. Homogenates were centrifuged at 10,000 RPM for 3 minutes at 4°C and pellets washed with cold PBS followed by resuspension in TSEI with Protease Inhibitor Cocktail. Samples were sonicated 12 times for 10 seconds and supernatants transferred to new tubes and incubated with rabbit IgG and anti-AhR or just rabbit IgG overnight at 4°C.  Protein A/G-Sepharose was added and samples incubated for 1.5h.  Pellets were washed for 10 minutes in buffer 1, buffer 2, LiCl buffer, and TE. Protein-DNA complexes were then eluted using TE and cross-links reversed by overnight incubation at 65°C followed by DNA purification. Libraries were prepared using the MicroPlex kit (Diagenode) according to manufacturer's instructions