Freedom™ DG44 Kit
Freedom™ DG44 Kit
Gibco™

Freedom™ DG44 Kit

Gibco™ Freedom™ DG44 Kit is an easy-to-use, beginning-to-end product, for cloning and expression of recombinant proteins in dihydrofolate reductase deficient了解更多信息
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货号数量
A13737011 kit
货号 A1373701
价格(CNY)
327,996.00
Each
添加至购物车
数量:
1 kit
价格(CNY)
327,996.00
Each
添加至购物车
Gibco™ Freedom™ DG44 Kit is an easy-to-use, beginning-to-end product, for cloning and expression of recombinant proteins in dihydrofolate reductase deficient (DHFR-) Chinese Hamster Ovary cells. The Gibco™ Freedom™ DG44 Kit includes components for transfection, expression, clone creation, and stable cell line selection, all in a conveniently packaged kit. The Gibco™ Freedom™ DG44 Kit also delivers a proven host cell line with simplified commercial licensing options.

• Flexible commercial licensing without the need for royalties
• Proven host used in approved products and a wide array of proteins
• Complete workflow that can take your gene of interest from transfection to lead clone typically in less than 6 months
• Convenient packaging, for simplified ordering and storage

Commercial Licensing that is Simple, Flexible and Without Royalties
Multiple companies provide CHO (Chinese Hamster Ovary) stable cell line development services and products that require annual fees during development of your therapeutic protein. In addition, royalties may be required on the sales of your therapeutic protein upon commercialization. Gibco™ Freedom™ Kits provide you with the tools that enable you or your CRO provider to develop your stable cell line for just the price of the kit itself. Furthermore, as you move towards commercial use, a flexible license structure is available without the burden of royalties and works to keep it as simple as a onetime payment (Fig 1). For more information regarding licensing, email us at outlicensing@lifetech.com.

Host Cell Background with Proven Results
DG44 cells have been a choice of scientists for decades for the expression of IgG and numerous blood proteins. Protein expression levels can vary significantly depending on the class of protein, however gram per liter levels of IgG expression has been demonstrated using these DG44 parental cells. So you can be confident that if your work calls for use of DG44 cells, they will deliver expected protein levels.

Complete, Fast, and Efficient Workflow
The Gibco™ Freedom™ DG44 Kit’s complete, proven, and fully integrated workflow allows you to move from transfection to lead clone typically in less then 6 months, helping to reduce hands on time and Fulltime Equivalent (FTE) requirements (Fig 2). We have implemented a single round of MTX (methotrexate) amplification in the protocol, which in our hands provided up to 120 fold titer improvement in less than 6 weeks (Fig 3). The entire workflow is performed in animal origin free (AOF) medium. The kit also offers the following advantages:

• Cloning of one or two genes that encode your protein(s) of interest
• High efficiency transfection of DNA into DG44 Cells (cGMP-banked)
• Included flash drive with loaded manual

Access to Gibco™ Freedom™ Kit Support
If any specific support is needed beyond the manual, please contact your account manager to obtain a Process Science Manager consultation. In addition, one of our support scientists may check in with you to ensure that your experience with the kit is as successful as possible. For further assistance, please email our support team at GibcoServices@thermofisher.com.
仅供科研使用。不可用于诊断程序。
规格
产品线Freedom™
数量1 kit
Unit SizeEach
内容与储存
CD DG44 Medium: 2°C to 8°C, protect from light
CD OptiCHO™ Medium: 2°C to 8°C, protect from light
OptiPRO™ SFM: 2°C to 8°C, protect from light
FreeStyle™ MAX Reagent: 2°C to 8°C, do not freeze
Pluronic™: F68 15°C to 30°C

DG44 (CHO) cGMP Banked suspension dhfr (dihydrofolate reductase) Negative Cells: liquid nitrogen
One Shot™ TOP10 Chemically Competent E. coli: –80°C
pcDNA 3.3 TOPO™ TA Vector Kit: –5°C to –20°C
pOptiVEC™ TOPO™ TA Vector Kit: –5°C to –20°C
Gentamicin™: –5°C to –20°C

常见问题解答 (FAQ)

在Freedom DG44试剂盒中,将蛋白亚基插入pcDNA3.3-TOPO TA载体或pOptiVEC-TOPO TA载体,会产生不同的后续效应吗?

我们推荐您在建立稳转细胞系之前,将每一蛋白亚基克隆分别克隆至两个载体中,以确定何种载体组合能够在瞬转条件下提供最高的蛋白产量。

在Freedom DG44试剂盒中,由于MTX扩增仅对pOptiVEC TOPO-TA载体所表达的蛋白亚基起作用,我应如何确认pcDNA3.3-TOPO TA载体所表达的蛋白亚基也实现了扩增?

尽管MTX扩增的机理尚不十分清楚,不过人们认为这两个载体会趋向整合于相同的基因组位点,因此也会实现同步扩增。

在Freedom DG44试剂盒中,pOptiVEC-TOPO TA载体包含了一个DHFR营养缺陷型筛选标志物,这样用户就可免除蛋白生产过程中的抗生素筛选步骤。不过,这对于包含新霉素筛选标志物的pCDNA3.3-TOPO TA载体而言是不可行的。能否在蛋白生产过程中加入G418成份?

在小规模的起始实验中,G418的存在不会对蛋白生产造成影响。不过,我们并不推荐在大规模蛋白生产过程中(即在生物反应器中)添加G418。

In the Freedom DG44 kit, does it matter which protein subunit is inserted into the pcDNA3.3-TOPO TA versus the pOptiVEC-TOPO TA vectors?

We recommend cloning each protein subunit into each of the vectors to determine which vector combination gives the highest protein yield in transient transfection before making the stable cell line.

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

In the Freedom DG44 kit, since MTX amplification would only work for the protein subunit expressed by the pOptiVEC TOPO-TA vector, how can I ensure that the protein subunit expressed by pcDNA3.3-TOPO TA is also being amplified?

Even though the exact mechanism for MTX amplification is not well understood, the thinking is that both vectors tend to integrate in the same genomic locus and hence would get amplified at the same time.

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