Cat.No. : CSC-RO0788 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-RO0788 |
| Description | Ba/F3-EML4-ALK-G1202R/L1198F cell line is engineered to stably overexpress human EML4-ALK fusion protein with G1202R/L1198F mutation. |
| 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 |
Alectinib has emerged as a potent treatment option, yet many patients experience relapse due to secondary mutations in the ALK gene, such as G1202R and I1171N. These mutations present a challenge, as they confer resistance to first-line therapies. In recent studies, it was demonstrated that several lorlatinib-resistant ALK-compound mutants, including those with G1202R mutations, exhibit higher resistance levels than their single-mutant counterparts. Importantly, some of these compound mutants respond favorably to earlier generation ALK tyrosine kinase inhibitors (TKIs), such as crizotinib and alectinib, suggesting potential therapeutic strategies for managing resistance in clinical settings.
Figure 1. The researchers employed ENU mutagenesis screening to identify ceritinib- or lorlatinib-resistant ALK compound mutations in Ba/F3 cells, aiming to uncover therapeutic strategies for overcoming resistance in ALK-mutated NSCLC. (Okada K, et al., 2019)
The Human EML4-ALK-G1202R/L1198F Stable Cell Line from Creative Biogene is designed for researchers exploring these complex mutations and their implications for treatment. This cell line provides a valuable tool for studying drug sensitivity and resistance mechanisms in ALK-mutated NSCLC.
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Using the Human EML4-ALK-G1202R/L1198F Stable Cell Line - Ba/F3 product, I can clearly feel the increase in experimental efficiency. This cell line provides us with a stable and reproducible experimental model, allowing us to rapidly observe changes in gene expression and drug response, greatly reducing the experimental cycle.
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