Expi293™ 诱导性表达系统试剂盒
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Expi293™ 诱导性表达系统试剂盒

The Expi293 Inducible Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian了解更多信息
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货号数量
A412751 个试剂盒
货号 A41275
价格(CNY)
-
数量:
1 个试剂盒
The Expi293 Inducible Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian 293 cells. The system kit is centered on engineered Expi293F cells that stably express the tetracycline repressor (TetR) protein, thus enabling tetracycline-regulated protein expression. The Expi293F Inducible cell line was established by the stable integration of the regulatory plasmid pcDNA6/TR, which encodes the TetR gene under the control of the human CMV promoter. This cell line is a powerful tool for tetracycline-inducible high-yield expression of recombinant proteins.

Expi293 Inducible Expression System Kit features include:
• Tetracycline-inducible protein expression with extremely low basal expression levels
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• pcDNA5/TO expression vector for tetracycline-regulated expression of the gene of interest
• Suspension, high-density culture of Expi293F Inducible cells in Expi293 Expression Medium
• High-efficiency transient transfection using ExpiFectamine 293 reagent in combination with specialized transfection enhancers

The components included in the Expi293 Inducible Expression System Kit are: one vial of frozen Expi293F Inducible cells, 1 L of Expi293 Expression Medium, one ExpiFectamine 293 transfection kit sufficient to transfect 1 L of culture, 100 mL Opti-Plex Complexation Buffer, one pcDNA5/TO Mammalian Expression Vector kit, and the pRABBIT IgG IRES-EmGFP positive control vector. All components of the Expi293 Inducible Expression System are also available for individual purchase.

仅供科研使用。不可用于诊断程序。
规格
细胞系Expi293F™
输送类型转染
适用于(应用)蛋白表达
关键功能蛋白质制备
反应次数1 kit
产品线Expi293
产品类型表达系统试剂盒
数量1 个试剂盒
运输条件经批准可在室温下或者湿冰或干冰上运输
细胞类型Expi293F
促进剂CMV/TO
样品类型DNA
Unit SizeEach
内容与储存
• Expi293F 诱导细胞(1 x 107 个细胞/小瓶),储存于汽相液氮中
• 1 L Expi293 表达培养基,在 2–8°C 下储存(避光储存)
• ExpiFectamine 293 转染试剂盒,用于 1 L 培养,在 2–8°C 下储存
• 100 mL Opti-Plex 综合缓冲液,在 2–8°C 下储存(避光)
• 20 µg pcDNA5/TO 哺乳动物表达载体,在 2–8°C 下储存或在 -20°C 下长期储存。
•pRABBIT IgG IRES-EmGFP 阳性对照载体,在 2–8°C 下储存或在 20°C 下长期储存

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。

Can I leave my Expi293F culture expressing for more than 7 days?

The growth and expression characteristics of Expi293F cells are such that, by 7 days post-transfection, the culture medium should be close to being spent and maximal protein expression should have already been achieved. Continued incubation will result in a large decrease in cell culture viability.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the difference between the Expi293F Inducible Cell Line and the T-REx -293 Cell Line?

The Expi293F Inducible Cell Line was created using the same technology as the T-REx -293 Cell Line. Both cell lines have been created by stable transfection with the pcDNA 6/TR vector; thus, they ubiquitously express the tetracycline repressor (TetR) protein. The parental cell line of Expi293F Inducible cells is Expi293F Cells (Cat. No. A14527). The key differences between these 2 cell lines are:
- Expi293F Inducible cells are grown in suspension and are adapted to high-density culture (> 15x10E6 cells/mL).
- Expi293F Inducible cells grow in Expi293 Expression System and perform equivalently to parental Expi293F cells, thus making them ideal for high-yield, inducible protein expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What method do you recommend for quantifying rabbit IgG expression from pRABBIT IgG IRES-EmGFP Positive Control Vector, Cat. No. A39243?

The formation of intact IgG molecules may be quantified using a sandwich ELISA designed to capture and detect rabbit IgG. Besides the rabbit IgG positive control vector, reagents, and consumables that are included in the kit, you will also need purified rabbit IgG to be used as a standard, F(ab')2-Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Cat. No. A10547), Protein A Coated Plates, Clear, 96-Well (Cat. No. 15130), TMB Substrate Kit (Cat. No. 34021), SuperBlock (TBS) Blocking Buffer (Cat. No. 37535), and PBS or TBS buffer for washes. There is an example procedure in our Protein A Coated Plates manual (https://tools.thermofisher.com/content/sfs/manuals/MAN0011310_Thermo Scientific_ProteinA_G_AG_Coat_96Well_UG.pdf). Please note that our R&D scientists determine titer values from crude cell culture supernatants using a Pall Life Sciences FortéBio Octet instrument equipped with a Protein A biosensor.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Do you offer a GFP-expressing mammalian expression vector that I can use as a control to monitor my transfection and expression with the Expi systems?

We offer pRABBIT IgG IRES-EmGFP Positive Control Vector, Cat. No. A39243, which you can use to monitor your transfection and expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.