ENCBS449OCN / cell line
Summary
- Status
- released
- Term name
- HepG2
- Term ID
- EFO:0001187
- Summary
- Homo sapiens HepG2 cell line genetically modified (insertion) using CRISPR targeting H. sapiens HDAC1
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-HDAC1-1
Genetic modifications
Accession | Category | Purpose | Method | Nucleic acid delivery method | Site |
|---|---|---|---|---|---|
| ENCGM880CRP | insertion | tagging | CRISPR |
|
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: HDAC1 Western (Lane 3). Predicted size was 58 kDa. The observed size was 70…
- Caption
- HDAC1 Western (Lane 3). Predicted size was 58 kDa. The observed size was 70 kDa, which is within 20% of an observed band of 68 kDa seen in https://www.proteinatlas.org/ENSG00000116478-HDAC1/antibody#western_blot. Each nuclear protein isolate (225 mcg - AHDC1; 209 mcg - HDAC1; 184 mcg - HMGN3; 297 mcg - SKI; 200 mcg - TIGD1; 248 mcg - ZFP82; 214 mcg - ZNF33A; 101 mcg - HepG2 Wild-Type; 291 mcg - SNAI1) 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. A HepG2 untagged nuclear isolate was included as a negative control, and a ZNF7-tagged nuclear isolate as a positive control. 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).
- Submitted by
- Scott Newberry
- Lab
- Richard Myers, HAIB
- Grant
- UM1HG009411