GSM2066432: HCC1954 LCC2 Input; 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
HCC1954
Primary Tissue
Breast
Site of Extraction
Soft Tissue
Tissue Diagnosis
Adenocarcinoma Ductal
Attributes by original data submitter
Sample
source_name
HCC1954 Cancer Cell Line
cell line source
HCC1954
source cell type
breast adenocarcinoma cell line
cell subtype
Latency Competent Cancer Cell (LCC) Derivative
chip antibody
none (Input)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were grown to 90–95% confluence, fixed with 1% formaldehyde for 15 min at room temperature followed by addition of glycine (1.25mM) for 10 minutes to stop cross linking. Nuclei were prepared and sonicated in lysis buffer (150 mM NaCl, 1% Triton X-100, 20 mM Tris pH 8.0, 0.1% SDS) using a Bioruptor to fragment chromatin to less than 500 base pairs (bp). Chromatin was pre-cleared with 35 μl Protein-G Dyna magnetic beads, and then immunoprecipitated with 5 μg of antibody over night at 4 °C. The chromatin antibody mix was incubated with 35 μl protein G-conjugated Dyna beads for 4 h at 4 °C, washed three times with wash buffer (1% Triton X-100, 50 mM Tris pH 8.0, 10% glycerol) with increasing concentrations of NaCl (150 mM, 300 mM and 400 mM), and two times with Tris-EDTA buffer. DNA was eluted in 1% SDS in Tris-EDTA buffer for 45 min at 37 °C, crosslinking was reversed overnight at 65 °C, and DNA was purified using Qiagen columns. For each ChIP, one fifth of the cells from 15cm plates were used.