GSM3307906: BIN-67 45D SMARCA4.K785R ChIP-Seq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
TFs and others
Cell type Class
Attributes by original data submitter
45D (Abcam, ab134942, GR3185597-1)
Sequenced DNA Library
cells were fixed in 1% formaldehyde at 37 degrees Celsius for 10 minutes, quenched in 2.5 M glycine, and snap frozen. Cells were thawed and lysed and the prepared nuclei was placed in a Covaris miliTube and sonicated for 20 minutes on a Covaris AFA Sonicator at standard settings (140 PIP, 5 W, 10% duty factor). The equivalent of 10 million cells were used in each immunoprecpitation. Three micrograms of the following antibodies were used: anti-SMARCA4 (Abcam ab110641); anti-DPF2 (Abcam); anti-SMARCC1 (homemade rabbit antibody raised against amino acids 998-1073 of human protein); anti-SS18 (Cell Signaling Technologies D6I4Z); anti-ARID2 (Cell Signaling Technologies D8D8U); anti-H3K4me1 (Abcam); anti-H3K27ac (Abcam). IP's were washed in 150 and 500 mM salt solution as well as a LiCl wash. Protein-DNA fragments were eluted using an SDS/DTT buffer and reverse crosslinked overnight. DNA was captured with SPRI beads (Agilent). Size selection was used to eliminate DNA too large for library prep. Rubicon library prep was used to generate libraries, and libraries were sequenced on Illumina NextSeq 500. ChIP-sequencing libraries were prepared with the Rubicon Thruplex DNAseq prep kit using standard protocols.