ENCBS734JXY / cell line

Summary

Status
released
Term name
HepG2
Summary
Homo sapiens HepG2 cell line genetically modified (insertion) using CRISPR targeting 3xFLAG-PBX2
Description
A standard Myers lab growth of HepG2, hepatocellular carcinoma
Product ID
HB-8065
Lot ID
n/a
Culture start date
2015-03-01
Culture harvest date
2015-03-18
Passage number
4

Attribution

ENCODE3 project
Lab
Richard Myers, HAIB
Award PI
Richard Myers, HAIB
Submitted by
Collin White
Source
ATCC
Project
ENCODE
Aliases
richard-myers:PBX2-FLAG-hepG2-1
Submitter comment
sonication date: 2015-07-22

Genetic modifications

Accession
Category
Purpose
Method
Site
ENCGM249WDXinsertiontaggingCRISPR

Donor information

Status
released
Accession
ENCDO000AAC
Aliases
encode:donor of HepG2, bradley-bernstein:Donor of HepG2
Donor external identifiers
Species
Homo sapiens
Life stage
child
Age
15 years
Sex
male
Health status
hepatocellular carcinoma
Ethnicity
Caucasian
References

Documents

growth protocol
Description: HepG2 culture conditions
Submitted by
Unknown User
Lab
Unknown Lab
immunoprecipitation followed by mass spectrometry
Caption excerpt: HepG2 whole cell lysate was immunoprecipitated using the primary antibody…
Attachment from submitter
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; F1804). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. The whole lane was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. Analysis of whole lane gel from HepG2: The sample was analyzed on a LTQ XL Linear Ion Trap Mass Spectrometer by LC-ESI-MS/MS. Peptides were identified using SEQUEST tandem mass spectral analysis with probability based matching at p < 0.05. SEQUEST results were reported with ProteinProphet protXML Viewer (TPP v4.4 JETSTREAM) and filtered for a minimum probability of 0.9. All protein hits that met these criteria were reported, including common contaminants. Fold enrichment for each protein reported was determined using a custom script based on the FC-B score calculation from the reference Mellacheruvu et al., 2013. The CRAPome: a contaminant repository for affinity purification mass spectrometry data. Nat. Methods. 10(8):730-736. Doi:10.1038/nmeth.2557. The target protein, HHEX, was identified as the 2nd enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
Submitted by
Keenan Graham
Lab
Richard Myers, HAIB