HK-ZFN-AURKB-mEGFP/ZFN-INCENP-mCherry 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/ZFN-INCENP-mCherry cell line, derived from HeLa Kyoto cells, is a specialized model used in cellular biology research. It has been genetically engineered to express Aurora B kinase (AURKB) tagged with monomeric enhanced green fluorescent protein (mEGFP) and Inner Centromere Protein (INCENP) tagged with mCherry. These modifications enable researchers to track the dynamics and interactions of these proteins during cell division. Aurora B kinase is essential for chromosome segregation and cytokinesis, while INCENP is a critical component of the Chromosomal Passenger Complex (CPC), coordinating mitotic progression. This dual fluorescent tagging provides a powerful tool for live-cell imaging, allowing detailed study of protein distribution during the cell cycle. The HK-ZFN-AURKB-mEGFP/ZFN-INCENP-mCherry cell line is valuable for researching mitotic regulation, chromosomal stability, and the mitotic checkpoint. The precision of zinc finger nucleases (ZFNs) used for genetic modifications ensures the accuracy of this model, making it ideal for high-fidelity studies in cancer biology and therapeutic development. |
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| Organism | Human |
| Tissue | Endocervix |
| Disease | Adenocarcinoma |
| Synonyms | HK-ZFN-AURKB-mEGFP,ZFN-INCENP-mCherry |
Characteristics
| Age | 30 years |
|---|---|
| Gender | Female |
| Ethnicity | African American |
| Morphology | Epithelial-like cells with mosaic stone shape |
| Growth properties | Adherent |
Regulatory Data
| Citation | HK-ZFN-AURKB-mEGFP/ZFN-INCENP-mCherry (Cytion catalog number 300270) |
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| Biosafety level | 1 |
| NCBI_TaxID | 9606 |
| CellosaurusAccession | CVCL_VL14 |
| Depositor | The Ellenberg Lab (EMBL) |
| GMO Status | GMO-S1: This HeLa Kyoto dual-color line contains ZFN-engineered AURKB-mEGFP and INCENP-mCherry constructs for chromosome passenger complex studies. 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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