ENCBS246HTK / cell line

Summary

Status
released
Term name
HepG2
Summary
Homo sapiens HepG2 cell line genetically modified (insertion) using CRISPR targeting 3xFLAG-TGIF2

Attribution

ENCODE3 project
Lab
Richard Myers, HAIB
Award PI
Richard Myers, HAIB
Submitted by
Collin White
Source
ATCC
Project
ENCODE
Aliases
richard-myers:TGIF-FLAG-hepG2-1

Genetic modifications

Accession
Category
Purpose
Method
Site
ENCGM034UISinsertiontaggingCRISPR

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

immunoprecipitation followed by mass spectrometry
Caption excerpt: HepG2 whole cell lysate was immunoprecipitated using the primary antibody…
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, TGIF2, was identified as the 2nd enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
Submitted by
Carrie Davis
Lab
Richard Myers, HAIB