Human EML4-ALK-G1202R/F1174L Stable Cell Line - BaF3

Name: Human EML4-ALK-G1202R/F1174L Stable Cell Line - BaF3
SKU: CSC-RO01236
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-RO01236

Host Cell : Ba/F3 Size : >1x10⁶ frozen cells/vial

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Cat. No.	CSC-RO01236
Description	This cell line is engineered to stably overexpress human EML4-ALK fusion protein with G1202R-F1174L mutations in ALK region. Expression of human EML4-ALK-G1202R-F1174L in this cell line has been analyzed by qPCR. Overexpression of human EML4-ALK-G1202R-F1174L render the BaF3 cells to be depended on the activation of EML4-ALK-G1202R-F1174L instead of IL-3 supplement. This cell line can grow in the medium without IL-3.
Product Type	Stable cell line constitutively expressing human kinase gene(s)
Target Gene	EML4-ALK-G1202R/F1174L
Gene Species	Homo sapiens (Human)
Host Cell	Ba/F3
Host Cell Species	Mus musculus (Mouse)
Applications	Drug screening and biological assays
Biosafety Level	2
Size	>1x10^6 frozen cells/vial, 1 mL
Stability	Stable in culture over a minimum of 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Growth Conditions	37 °C, 5% CO₂
Storage	Liquid nitrogen
Shipping	Dry ice

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.
Freeze Medium	Frozen with 70% medium, 20% FBS, 10% DMSO
Growth Properties	Suspension, round
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

CSC-RO01236-Human EML4-ALK-G1202R/F1174L Stable Cell Line - BaF3-Datasheet

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EML4 gene is a member of the echinoderm microtubule associated protein-like family. The encoded WD-repeat protein may be involved in microtubule formation. ALK gene encodes a receptor tyrosine kinase, which belongs to the insulin receptor superfamily. ALK protein comprises an extracellular domain, an hydrophobic stretch corresponding to a single pass transmembrane region, and an intracellular kinase domain. It plays an important role in the development of the brain and exerts its effects on specific neurons in the nervous system. This gene has been found to be rearranged, mutated, or amplified in a series of tumours including anaplastic large cell lymphomas, neuroblastoma, and non-small cell lung cancer. Abnormal fusion of parts of EML4 gene with portions of the anaplastic lymphoma receptor tyrosine kinase gene, which generates EML4-ALK fusion transcripts, is one of the primary mutations associated with non-small cell lung cancer.

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