Expi293F™ 细胞
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Expi293F™ 细胞
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Expi293F™ 细胞

Expi293F™ 人细胞来源于 293 细胞系,是 Expi293™ 表达系统的核心组分
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货号数量
A145286 x 1 mL
A145271 管
A14527CN1 管
货号 A14528
价格(CNY)
-
数量:
6 x 1 mL

Expi293F human cells are derived from the 293 cell line and are a core component of the Expi293 Expression System. They are maintained in suspension culture and will grow to high density in Expi293 Expression Medium. Expi293F cells are highly transfectable and generate superior protein yields compared to standard 293 cell lines in transient protein expression.

Expi293F Cells attributes include:

  • Suspension culture cells pre-adapted to Expi293 Expression Medium
  • Demonstrated higher protein expression levels than other 293 cells adapted to Expi293 Expression Medium
  • Exhibit more rapid cell growth and higher culture viabilities than standard suspension-adapted 293 cells
  • Consistent transient expression performance of Expi293F cells for over 30 passages beyond thaw
  • Also available from a cGMP bank (Cat. No. 100044202)

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in Expi293 Expression Medium and 10% DMSO. The cells should be thawed directly into Expi293 Expression Medium. The six-vial multipack eliminates the need to expand and rebank the cells in the lab.

Cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable from volumes of <1 mL in multiwell plates to >10 L in bioreactors. Protein harvest can be performed at 2–7 days post-transfection, depending on the protein expressed. For expression of recombinant antibodies, harvest is typically at 5–7 days post-transfection. There is no need for culture medium supplementation or batch feeding during expression.

仅供科研使用。不可用于诊断程序。
规格
推荐培养基Expi293™ 表达培养基
池数量1 x 107
产品线Expi293
产品类型293-F 细胞
数量6 x 1 mL
细胞系Expi293F™
细胞类型肾脏(胚胎)
培养类型悬浮液细胞培养
形式冻存
种属
Unit SizeEach
内容与储存
1 x 107 个细胞/mL,在液氮中储存。

常见问题解答 (FAQ)

我能让Expi293F培养细胞表达蛋白长达7天以上吗?

Expi293F细胞的生长和表达属性决定了在转染7天之后,培养基将临近耗竭,并应该已经达到最高蛋白表达量。继续孵育将导致培养细胞活力的显著下降。

在使用Expi293表达系统的过程中,我应等待多少天后收获蛋白?

最佳的表达时间随不同的目的蛋白而发生变化,但大多数测试蛋白的最佳表达时间窗为3-7天。由于本系统具有良好的扩展性,我们推荐您可在放大扩种之前进行一个小规模测试实验,来决定何时收获目的蛋白。

若使我的Expi293F细胞生长至比推荐指标更高的细胞密度,它们还能用吗?

尽管Expi293培养基能够支持更高的细胞密度,但我们并不推荐以超出5–6 x 10e6个细胞/毫升的密度来培养Expi293细胞,因为这会降低后续的转染和蛋白表达效率。在更高的细胞密度时,遇到培养活力降低临界点的可能性也会增加。如果您培养的细胞超出5–6 x 10e6个细胞/毫升,请按照手册中的描述传代一至两次,监控它们的活力和生长速率。使用蛋白表达对照IgG或得率已知的表达载体来测试转染效果,从而确定细胞表达性能是否受到影响。

我能在Expi293表达系统中使用另外一种293细胞系吗?

大多数293细胞系均能适应从传统含血清或其它无血清培养基向Expi293表达培养基中的切换按照手册中所描述的直接切换法或序贯切换法来操作,大多数293细胞系均能适应从传统含血清或其它无血清培养基向Expi293表达培养基中的切换。 注意:切换后的细胞可能无法达到与Expi293F细胞同等的高水平表达。

你们推荐在Expi293表达系统中使用何种载体?

我们推荐使用pcDNA 3.4-TOPO TA载体(货号A14697)。这一载体含有天然的全长CMV启动子和克隆位点下游的WPRE(旱獭转录后调控元件),这两个元件均有利于基因的高水平表达(表达比pcDNA3.3 TOPO载体高大约2-3倍,而后者又比标准的pcDNA载体高2-5倍)。当然,实际表达水平还要视具体蛋白而定。