| Field | Specification |
|---|---|
| Organism | |
| Applications | |
| Tissue | |
| Disease | Hepatocellular carcinoma |
| Age | |
| Sex | |
| Ethnicity | |
| Growth properties | |
| Biosafety level | |
| Product format | Frozen |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Overview
This product is a stable CRISPR knockout cell line in which MCFD2 has been disrupted in the (Human (H. sapiens)) background using CRISPR-Cas9 genome editing. It provides an isogenic model for loss-of-function studies of MCFD2 in liver tissue context. Cells are supplied frozen (1×10⁶ cells / 1.0 ml, BSL-II) and grow as adherent, epithelial-like monolayers.
CRISPR Knockout Design
- Target gene: MCFD2
- Knockout strategy: CRISPR-Cas9–mediated frameshift-inducing INDEL generation; confirmed by sanger sequencing of frameshift-inducing indels
- Design notes: This HepG2 cell line features a stable knockout of MCFD2 and is applicable for studying the ER-to-Golgi transport mechanism of blood coagulation factors and proteins such as α1-antitrypsin.
- Parental cell line: — Human (H. sapiens), liver origin
Gene Background
MCFD2 is a protein-coding gene. Its encoded protein participates in cellular processes relevant to the biological pathways associated with liver biology. For detailed gene function, pathway context, and disease associations, refer to the NCBI Gene and UniProt entries for MCFD2.
Cell Culture Specifications
- Culture medium
- PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. Eagle's Minimal Essential Medium (EMEM) (TM511) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate
- Growth properties
- Adherent, epithelial-like
- Tissue origin
- Liver
- Organism
- Human (H. sapiens)
- Biosafety level
- BSL-II
- Format
- Frozen; 1×10⁶ cells / 1.0 ml
Quality is assessed by: Confirmed by Sanger Sequencing of frameshift-inducing INDELs.
Research Applications
- Validate MCFD2 knockout by Western blot or qPCR versus the isogenic parental line.
- Investigate loss-of-function phenotypes: proliferation, viability, morphology, or pathway activity changes.
- Screen drug candidates targeting pathways regulated by MCFD2 using this KO as an isogenic control.
- Perform rescue experiments by re-expressing wild-type MCFD2 in the knockout background.
- Conduct transcriptomic or proteomic profiling comparing KO versus parental cells.
Important Notes
- Add selection antibiotic to culture medium only after the first passage post-thaw to allow cells to recover.
- Cell viability upon thaw is warranted for 30 days following shipment when handled per abm guidelines.
- For laboratory research use only. Not intended for diagnostic, therapeutic, or clinical applications.
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
