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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 deficientRead more
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Catalog NumberQuantity
A13737011 kit
Catalog number A1373701
Price (EUR)
17 060,00
Each
Estimated availability date 16-Jul-2025
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Quantity:
1 kit
Recurring order eligible. Learn more »
Price (EUR)
17 060,00
Each
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Ask our AI about this Product
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.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product LineFreedom™
Quantity1 kit
Unit SizeEach
Contents & Storage
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
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Frequently asked questions (FAQs)

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.

In the Freedom DG44 kit, the pOptiVEC-TOPO TA vector contains a DHFR auxotrophic selection marker so that one can avoid using antibiotic selection during protein production. However, this is not possible with the pcDNA3.3-TOPO TA vector as it contains a neomycin selection marker. Is it okay to have G418 present during protein production?

Using G418 should not pose a problem for low-scale protein production for initial studies. However, we do not recommend adding G418 during large-scale protein production (i.e., in a bioreactor).

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

Can MTX be used at concentrations higher than 4 µM to increase protein yield using the OptiCHO Express Kit?

There are a couple of concerns associated with high MTX concentrations. One is the stability of the cell line generated using high MTX concentrations. The other is that it may be amplifying the DFHR gene without amplifying the gene of interest along with it stoichiometrically. We recommend the more conservative approach of 4 µM MTX for amplification.

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

How can I learn more about commercial licensing of Freedom kits, ExpiCHO-S Cells (cGMP banked), CHO-S Cells (cGMP banked), and CHO DG44 Cells (cGMP banked)?

To inquire about a Commercial Production or Service License, please email: outlicensing@thermofisher.com.

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

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Notice to Purchaser: The purchase of this product does not convey to the purchaser any license to any patents or patent applications related thereto. Each purchaser of this product should consider whether additional rights are required for the purchaser's particular application under the above patents.

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