HK-ZFN-AURKB-mEGFP Cells
USD$800.00*
Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 106 cells for adherent lines or 5 × 106 cells for suspension lines (refer to the batch CoA for details).
General information
| Description | The HK-ZFN-AURKB-mEGFP cell line is a genetically engineered human cell model designed to express the AURKB (Aurora Kinase B) protein fused with mEGFP (monomeric Enhanced Green Fluorescent Protein) using Zinc Finger Nuclease (ZFN) technology. AURKB is a serine/threonine kinase that plays a crucial role in mitotic chromosome segregation, cytokinesis, and the regulation of the mitotic spindle checkpoint. The fusion with mEGFP allows for real-time visualization of AURKB activity and localization within the cell, facilitating detailed studies of its dynamic behavior during cell division. This cell line serves as a powerful tool for researchers investigating the molecular mechanisms of mitosis and the specific functions of AURKB. The incorporation of mEGFP enables fluorescence-based assays and live-cell imaging, providing insights into the spatiotemporal distribution of AURKB. The use of ZFN technology ensures precise genomic integration, maintaining the fidelity of AURKB expression. This model is particularly valuable in cancer research, where AURKB is often overexpressed and linked to tumorigenesis, making it a potential target for therapeutic interventions. |
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| Organism | Human |
| Tissue | Endocervix |
| Disease | Adenocarcinoma |
Characteristics
| Age | 30 years |
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| Gender | Female |
| Ethnicity | African American |
| Morphology | Epithelial-like cells with mosaic stone shape |
| Growth properties | Adherent |
Regulatory Data
| Citation | HK-ZFN-AURKB-mEGFP (Cytion catalog number 300173) |
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| Biosafety level | 1 |
| NCBI_TaxID | 9606 |
| CellosaurusAccession | CVCL_VL13 |
| Depositor | The Ellenberg Lab (EMBL) |
| GMO Status | GMO-S1: This HeLa Kyoto line contains a ZFN-integrated mEGFP fusion at the endogenous AURKB locus for mitotic kinase imaging. This classification applies only within Germany and may differ elsewhere. |
Biomolecular Data
| Products | EGFP (Enhanced Green Fluorescent Protein) |
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Handling
| Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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| Supplements | Supplement the medium with 10% FBS |
| Dissociation Reagent | Accutase |
| Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
| Fluid renewal | 2 to 3 times per week |
| Freeze medium | As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. |
| Thawing and Culturing Cells |
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| Incubation Atmosphere | 37°C, 5% CO2, humidified atmosphere. |
| Shipping Conditions | Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage. |
| Storage Conditions | For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen. |
Quality Control & Molecular Analysis
| Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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