CD 293 TGE Medium, 1X, Liquid
产品描述（Description）
CD 293 TGE培养基是一种有专利的、化学定义的、无动物来源的培养基，专门用于293细胞的高密度悬浮培养和转染。质粒DNA在适应于该培养基的HEK293细胞中的瞬时表达可以获得高体积产率的靶蛋白。建议将293 Expression MAX-1和Feed X Supplement与BPM BPfectin一起使用，以获得更高的蛋白质表达率。
CD 293 TGE培养基专为以下用途配制：
1. 悬浮液HEK293（293T、293EBNA、293F）瞬时表达
2. HEK 293稳定蛋白表达细胞系的悬浮培养
3. HEK293细胞在生物反应器中的大规模高密度生长
产品特性（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
产品参数（General Specification）
操作流程（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.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).
⑥ Incubate the complexes at RT for 10 min before transfection.
⑦ Add DNA-BPfectin complexes to cells at 5% volume ratio mix well gently.
⑧ 24 hours post transfection, add Feed X Supplement (Catalog # CF-1116) to culture at 10% volume ratio (e.g. 1 mL to 10mL culture).
⑨ Harvest for purification typically 7 days post transfection or when cell viability drops below 55%.
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