SAMD00022899
External Id
Hokkaido University Department of Biochemistry, Hokkaido University Graduate School of Medicine
INSDC center name
2016-10-06T16:21:24Z
INSDC first public
2016-10-06T03:21:24Z
INSDC last update
DRS034280
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
DRS034280
SRA accession
HEK293T
http://www.ebi.ac.uk/efo/EFO_0001184
cell line
Cells from one 10 cm dish (~1 x 107) of HEK293T cells grown to 70-80% confluence were used for each immunoprecipitation. The cells were cross-linked with 2 mM DSG Crosslinker in PBS for 30 min and then 1% formaldehyde in PBS for 20 min at room temperature. Then the cells were resuspended and lysed in lysis buffer (0.5% SDS, 10mM EDTA, 150 mM NaCl, 50 mM Tris-HCl pH 8.0), and they were sonicated with a Bioruptor Sonicator (Diagenode, Denville, NJ) for 30 times for 30 seconds at the maximum power setting to generate DNA fragments of ~150-500 bps. Sonicated chromatin was incubated at 4C overnight with 10 ug of anti- ICE1 (raised against bacterially expressed, His-tagged recombinant proteins of ICE1 1865-2266 aa). Crosslinking was reversed by overnight incubation at 65C. Immunoprecipitated DNA were treated with RNase A and Proteinase K by incubation at 45C. DNA was purified using the QIAquick MinElute PCR purification kit (28006, Qiagen).
description
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
KIAA0947
sample name
HEK293T_ICE1_ChIPseq
title