CD 293 TGE Medium, 1X, Liquid

Description

CD 293 TGE Medium is a proprietary, chemically defined, animal origin-free medium specifically developed for the high-density, suspension culture and transfection of 293 cells. Transient expression of plasmid DNA in HEK293 cells adapted in this medium can get high volumetric productivity of target proteins. Used with BPM BPfectin, 293 Expression MAX-1 and Feed X Supplement together is recommended for higher yield of protein expression.

CD 293 TGE Medium is specifically formulated for use with:

1. Suspension HEK293(293T, 293EBNA, 293F) transient expression
2. Suspension culture of HEK 293 stable cell line for protein expression
3. Large-scale, high-density growth of HEK293 cells in bioreactors

Product Features

1. Support suspension HEK 293(293T, 293EBNA, 293F) transient expression
2. >200% cell growth and expression performance compared competitive alternatives
3. Prepared ready-to-use, with no supplementation required
4. Protein free, animal component free, chemically define and regulatory friendly
5. Extensive support by cell culture expert

General Specification

Usage Protocol

1. Before transfection, passage 293 cells with CD 293 TGE Medium for at least 3 round from an adaptation procedure, seeding density greater than 0.5 ×106 cells/ml, cell viability >95%. Culture at 150-180 rpm depending on your orbital shaker design and single cell distribution status.
2. Seed 293 cells at 0.5 ×106 cells/ml and transfer to pre-warmed fresh medium, subsequently grow the cells to 1.5 to 2.2 ×106 cells/ml.
3. The day before transfection, dilute the culture with fresh medium to 0.8 to 1.0 ×106 cells/ml and cell density at transfection should range from 1.5 × 106 to 2.0 × 106 cells/ml, provided the doubling time is 24 h and viability is greater than 95%.
4. On the day of transfection, using sterile 150 mM NaCl solution to dilute DNA plasmid and add 293 Expression MAX-1 (Catalog # EXP-711) according to the instruction.
5. Add BPfectin (Catalog # TF-1157) at ratio of 3:1 (BPfectin : DNA; volume/weight) to DNA solution mix well with vortex for 3 sec.
6. Note: DNA dosage for transfection is 1 ug per 1 million cells in culture, and volume of DNA-BPfectin complexes is 5%(v/v) of culture. (e.g. 500 μL to 10 mL culture).
7. Incubate the complexes at RT for 10 min before transfection.
8. Add DNA-BPfectin complexes to cells at 5% volume ratio mix well gently.
9. 24 hours post transfection, add Feed X Supplement (Catalog # CF-1116) to culture at 10% volume ratio (e.g. 1 mL to 10mL culture).
10. Harvest for purification typically 7 days post transfection or when cell viability drops below 55%.

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