GSM4462229: SUM159 ZNF446 input rep1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Breast
Cell type
SUM 159PT
Primary Tissue
Breast
Tissue Diagnosis
Anaplastic Carcinoma
Attributes by original data submitter
Sample
source_name
Human triple-negative breast cancer cells
cell line
SUM159
cell type
triple-negative breast cancer
chip antibody
input
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were crosslinked with 1% formaldehyde for 15 min at 37°C. Crosslinking was quenched with 0.125 M glycine for 5 min at 4°C, followed by multiple washes with cold PBS. Nuclei were harvested via hypotonic lysis and subjected to sonication in a lysis buffer containing 1% SDS. Sheared chromatin lysates were then pre-cleared with Protein A/G agarose beads for 1 hour, followed by immunoprecipitation with appropriate antibody overnight at 4°C. Protein A/G agarose beads blocked with BSA were added to lysates for 2 hours the next day, followed by extensive washes. 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).