Cat.No. : CSC-RO0326 Host Cell: Ba/F3
Size: >1x10^6 frozen cells/vial, 1 mL Stability: Stable in culture over a minimum of 10 passages
| Cat. No. | CSC-RO0326 |
| Description | This cell line is engineered to stably overexpress exogenous EML4-ALK fusion gene with F1174C and D1203N mutations in the ALK part. |
| Target Gene | EML4-ALK |
| Gene Species | Homo sapiens (Human) |
| Host Cell | Ba/F3 |
| Host Cell Species | Mus musculus (Mouse) |
| Stability | Stable in culture over a minimum of 10 passages |
| Application | Drug screening and biological assays |
| Growth Conditions | 37 °C, 5% CO2 |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Size | >1x10^6 frozen cells/vial, 1 mL |
| Biosafety Level | 2 |
| Thawing & Subculturing Instructions | 1. Thaw cells by gently swirling in a 37°C water bath. To limit contamination, do not submerge the O-ring and cap. 2. When cells are ~70% thawed (~1 min), transfer the vial into a biosafety cabinet, and wipe the surface with 70% ethanol. Allow tube to dry completely. 3. Transfer the cells gently into a 15 mL conical tube containing 10 mL of pre-warmed culture medium (without antibiotic selection marker). Centrifuge cells at ~125 x g for 5~7 min. 4. Remove supernatant without disturbing the pellet, and resuspend cells in 1 mL culture medium (without antibiotic selection marker). Transfer cells to a 6-well plate containing ~2 mL pre-warmed growth medium (without antibiotic selection marker) or a T25 flask containing 5 mL pre-warmed culture medium (without antibiotic selection marker). 5. Incubate the culture at 37°C with 5% CO2. 6. Subculture: split saturated culture 1:4 ~ 1:6 every 3 days; seed out at about 1~3 x 10^5 cells/mL. |
| Growth Properties | Suspension, round |
| Freeze Medium | Frozen with 70% medium, 20% FBS, 10% DMSO |
| Freezing Instructions | Cells are recommended to generate additional frozen stocks at early passages. Frozen stocks should be preserved in a designated cryopreservation medium or in 70% RPMI 1640 + 20% FBS + 10% DMSO (without antibiotic selection marker). 1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO, without antibiotic selection marker) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x10^6 cells/ml in chilled freezing medium. 6. Aliquot 1 ml of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
A: The EML4-ALK-D1203N-F1174C gene represents a specific variant of the EML4-ALK fusion gene, which is found in some cases of non-small cell lung cancer (NSCLC). The D1203N and F1174C mutations occur within the ALK (anaplastic lymphoma kinase) portion of the fusion protein, altering its structure and function. This variant is known to contribute to the development of aggressive lung cancer and is a target for targeted therapies.
A: The D1203N and F1174C mutations in the EML4-ALK fusion protein affect ALK function by altering its kinase domain. These changes can lead to increased kinase activity, which in turn promotes uncontrolled cell growth and survival. This hyperactivity is a key driver of the oncogenic process in some NSCLC cases.
A: Targeting the EML4-ALK-D1203N-F1174C fusion gene has significant therapeutic implications. Inhibitors of ALK kinase activity, such as crizotinib (Xalkori) and newer ALK inhibitors, can be effective in treating NSCLC with this specific fusion variant. These targeted therapies can lead to tumor regression and improve patient outcomes.
A: The EML4-ALK-D1203N-F1174C fusion gene is detected through molecular diagnostic techniques such as reverse transcription polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH), and next-generation sequencing (NGS). These methods can identify the specific fusion event and the presence of the D1203N and F1174C mutations, allowing for accurate diagnosis and treatment planning for patients with NSCLC and this fusion variant.
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The Human ETV6-SYK Stable Cell Line - BaF3 has been widely cited in scientific literature, indicating its significant contribution to research on ETV6-SYK fusion kinase and its role in hematological malignancies.
Offering a range of cell lines, the Human ETV6-SYK Stable Cell Line - BaF3 adds to the diversity of research tools available for studying hematological cancers, enabling researchers to explore various aspects of the disease.
Researchers appreciate the provider's willingness to offer customization services for the Human ETV6-SYK Stable Cell Line - BaF3, allowing for tailored experiments that meet specific research needs and objectives.
The Human ETV6-SYK Stable Cell Line - BaF3 undergoes rigorous quality control measures, ensuring that researchers receive a reliable and high-quality product that consistently performs in experiments.
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