ENCBS073YLW / cell line

Summary

Status
released
Term name
HepG2
Summary
Homo sapiens HepG2 cell line genetically modified (insertion) using CRISPR targeting H. sapiens FOXC1

Attribution

ENCODE4 project
Lab
Richard Myers, HAIB
Award PI
Richard Myers, HAIB
Submitted by
Scott Newberry
Source
Richard Myers
Project
ENCODE
External resources
Aliases
richard-myers:HepG2-3xFLAG-FOXC1-1

Genetic modifications

Accession
Category
Purpose
Method
Nucleic acid delivery method
Site
ENCGM608NLVinsertiontaggingCRISPR

Donor information

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

Documents

immunoblot
Caption excerpt: FOXC1 Western (Lane 3). Each nuclear protein isolate (134 mcg - ARID4A; 177…
Caption
FOXC1 Western (Lane 3). Each nuclear protein isolate (134 mcg - ARID4A; 177 mcg - FOXC1; 300 mcg - NCoA1; 176 mcg - ZBTB12; 201 mcg - ZMYM4; 295 mcg - ZNF30; and 150 mcg - ZNF827) was standardized in a solution containing a volume of 2% Halt Protease and Phosphatase Inhibitor Single-Use Cocktail Mixture (Thermo Fisher Scientific), NuPage Sample Reducing Agent 10X, and NuPage LDS Sample Buffer 4X (Thermo Fisher Scientific). After heating the solution for 15 minutes at 90C followed by cooling on ice, the protein samples were loaded onto a NuPage 4-12% Bis-Tris gel (Thermo Fisher Scientific) and separated using a PowerEase 90W system (Thermo Fisher Scientific) running at 150 V for 1 hour. The protein bands were transferred to a nitrocellulose membrane using the Invitrogen iBlot 2 System (Thermo Fisher Scientific), and blocked overnight at 4C in 5% milk solution with gentle rocking. The membrane was treated with a 1:5000 dilution of monoclonal M2-Peroxidase-conjugated ANTI-FLAG antibody (diluted in 5% BSA solution) (Sigma-Aldrich; cat# A8592) for 1 hour. Following four 5-minute washes with 1X TBST, visualization was attained with the Super Signal West Femto solution kit (Thermo Fisher Scientific) and a MyECL Imager (Thermo Fisher Scientific). The second western blot image depicts a negative control prepared with HepG2 nuclear lysate (Lane 2).
Submitted by
Scott Newberry
Lab
Richard Myers, HAIB