U2OS-CRISPR-NUP96-SNAP Cells
USUSD$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).
一般資訊
| 說明 | The U-2 OS-CRISPR-NUP96-SNAP is a genetically modified osteosarcoma cell line derived from the parent U-2 OS human cell line. This cell line has been engineered through CRISPR/Cas9-mediated genome editing to incorporate a SNAP-tag at the NUP96 gene, enabling the visualization and study of nuclear pore complex dynamics. Nuclear pore complexes (NPCs) are crucial for the regulation of nucleocytoplasmic transport, and NUP96 is a significant component of the NPC, playing a pivotal role in its structural integrity and function. In U-2 OS-CRISPR-NUP96-SNAP clone no.33, the integration of the SNAP-tag at the NUP96 locus allows for specific and covalent attachment of fluorescent substrates or other chemical probes that can be used for live-cell imaging and other biochemical assays. This feature makes it an invaluable tool for investigating the molecular dynamics of nucleocytoplasmic transport, understanding NPC-related pathologies, and screening for therapeutic compounds that affect NPC function. The cell line also retains the characteristics of the parental U-2 OS line, which includes a high level of genetic stability and ease of culture, making it suitable for high-throughput screening and extended studies in cell biology. Due to the specificity of the modification at the NUP96 gene, U-2 OS-CRISPR-NUP96-SNAP clone no.33 provides a unique model for the detailed study of NPC components in the context of cellular function and dysfunction. Researchers can exploit the SNAP-tag system to selectively and rapidly label NUP96, facilitating real-time visualization of NPC dynamics under physiological and pathological conditions. This specific clone can serve as a robust platform for both basic research and applied biomedical studies, contributing significantly to the fields of cellular biology, genetics, and oncology. |
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| 生物體 | Human |
| 組織 | Bone |
| 疾病 | Osteosarcoma |
特徵
| 年齡 | 15 years |
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| 性別 | Female |
| 族裔 | Caucasian |
| 生長特性 | Adherent |
監管數據
| 引用 | U-2 OS-CRISPR-NUP96-SNAP (Cytion catalog number 300444) |
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| 生物安全等級 | 1 |
| NCBI_TaxID | 9606 |
| Cellosaurus 編號 | CVCL_B7FL |
| 存款人 | The Ellenberg Lab (EMBL) |
| 轉基因狀態 | GMO-S1: This human osteosarcoma cell line (U2OS-CRISPR-NUP96-SNAP, clone 33) contains a CRISPR-engineered NUP96-SNAP fusion facilitating SNAP-tag chemical labeling of nuclear pores. The modification is stably integrated. This classification applies only within Germany and may differ elsewhere. |
生物分子資料
| 蛋白質表達 | NUP96-SNAP (nuclear pore complex protein 96, SNAP-tag) |
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處理方式
| 培養基 | McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a) |
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| 營養補充品 | Supplement the medium with 10% FBS, 3.0 g/L Glucose, stable Glutamine, 2.0 mM Sodium pyruvate, 2.2 g/L NaHCO3, 1% NEAA |
| 解離試劑 | Accutase |
| 傳代培養 | 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. |
| 播種密度 | 1 x 104 cells/cm2 |
| 流體更新 | 2 to 3 times per week |
| 冷凍培養基 | 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. |
| 細胞解凍與培養 |
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| 培養環境 | 37°C, 5% CO2, humidified atmosphere. |
| 運送條款 | 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. |
| 儲存條件 | 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. |
品質控制與分子分析
| 不孕症 | 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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分析證明書 (CoA)
| 批次編號 | 證書類型 | 日期 | 型號 |
|---|---|---|---|
| 300444-040724 | 分析證明書 | 23. May. 2025 | 300444 |