FreeStyle™ 293 Expression Medium
FreeStyle™ 293 Expression Medium
Gibco™

FreeStyle™ 293 Expression Medium

An animal origin-free, serum-free, protein-free formulation, allowing for easier purification
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Catalog NumberQuantity
123380181 L
123380266 x 1 L
1233800110 L
1233800220 L
Catalog number 12338018
Price (USD)
170.00
Each
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Quantity:
1 L
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Price (USD)
170.00
Each
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FreeStyle™ 293 Expression Medium is an animal origin–free, chemically defined, protein-free medium specifically developed to support the growth and transfection of 293-F cells in suspension without adaptation. The medium is complete and ready-to-use, with no supplementation required.

Features of Gibco™ FreeStyle™ 293 Expression Medium include:

  • Complete and ready-to-use format containing GlutaMAX™ supplement
  • Ability to support suspension growth and transfection of 293-F cells
  • Animal origin–free, protein-free and chemically defined formulation
  • Easy use with the FreeStyle™ 293 Expression systems
  • Scalability in spinner flasks and bioreactors

Complete and ready-to-use format containing GlutaMAX™ supplement

Gibco™ FreeStyle™ 293 Expression Medium is ready to use. It does not require the addition of serum, glutamine, or surfactants. Gibco™ FreeStyle™ 293 Expression Medium contains GlutaMAX™ supplement to minimize toxic ammonia build-up and improve cell viability and growth in an easy-to-use format.

Ability to support suspension growth and transfection of 293-F cells

Gibco™ 293-F, 293-H, and FreeStyle™ 293-F cells in suspension can be cultured directly into Gibco™ FreeStyle™ 293 Expression Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into Gibco™ FreeStyle™ 293 Expression Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations. Cells cultured in Gibco™ FreeStyle™ 293 Expression Medium can be transfected using lipid based transfection reagents, such as 293fectin™ reagent (Cat. No. 12347019) and Freestyle™ MAX reagent (16447500).

Animal origin–free, protein-free and chemically defined formulation

Gibco™ FreeStyle™ 293 Expression Medium is animal origin–free, protein-free and chemically defined, allowing for easier purification of your protein of interest. Gibco™ chemically defined media contain no proteins, hydrolysates, or components of unknown composition.

Easy use as part of the FreeStyle™ 293 Expression System and FreeStyle™ MAX Expression System for protein production

The FreeStyle™ 293 Expression System (Cat. No. K900001) is a complete, suspension cell culture system for generating large amounts of mammalian recombinant protein (see figure). The FreeStyle™ 293 Expression System combines Gibco™ FreeStyle™ 293 Expression Medium with 293fectin™ reagent, a cationic-lipid formulation designed to transfect suspension FreeStyle™ 293-F cells at high efficiency. The FreeStyle™ MAX Expression System (K900010) is a breakthrough technology for rapid and high-yield mammalian protein production. The FreeStyle™ MAX Expression System combines Gibco™ FreeStyle™ Media, FreeStyle™ MAX Reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293-F cells.

Scalability in spinner flasks and bioreactors

Protein production using Gibco™ FreeStyle™ 293 Expression Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement Gibco™ FreeStyle™ 293 Expression Medium with additional Pluronic™ F-68 to avoid sheer stress in the culture.

Universal bag packaging

Gibco™ universal bags offer flexible packaging that allows you to integrate a single-use, disposable large volume media container with a wide variety of automated cell culture instruments, disposable bioreactor systems, and sterile connection processes, minimizing the need for costly customization.

Product use

Customers using Gibco™ FreeStyle™ 293 Expression Medium in a manufacturing process, who have a submission with the FDA, may request a letter of authorization from us to reference our Type II Drug Master File (DMF).

cGMP manufacturing and quality system

Gibco™ FreeStyle™ 293 Expression Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.

For Research Use or Further Manufacturing. Not for diagnostic use or direct administration into humans or animals.
Specifications
Cell LineFreeStyle™ 293-F Cells
ClassificationAnimal Origin-free, Chemically-defined, Protein-free, Serum-free
Culture TypeSuspension Cell Culture
FormLiquid
Product TypeExpression Medium
Serum LevelSerum-free
Green FeaturesSustainable packaging
Product LineFreestyle™
Quantity1 L
Shipping ConditionRoom Temperature
Unit SizeEach
Contents & Storage
Storage conditions: 2°C to 8°C. Protect from light
Shipping conditions: Ambient
Shelf life: 12 months from date of manufacture

Frequently asked questions (FAQs)

Can I load the FreeStyle 293 Expression Medium or FreeStyle CHO Expression Medium directly onto a nickel column for His-tagged protein purification?

With FreeStyle 293 Expression Medium, we would recommend harvesting the cells a little earlier to reduce any nonspecific binding by host cell proteins and then filtering the culture supernatant through a 0.2 or 0.1 µM membrane before loading onto the column. On the other hand, FreeStyle CHO Expression Medium contains EDTA that will strip the nickel column. The medium can be subjected to dialysis or buffer exchange prior to loading on the column, or the EDTA can be chelated by adding 1-3 mg/L NiSO4 or NiCl to the supernatant. Iron or cobalt salts can also be used.

Can FreeStyle 293-F cells grown in FreeStyle 293 expression medium be transfected with ExpiFectamine293 Reagent along with enhancers? Is so, will the transfection efficiency be higher than that with FreeStyle Max Reagent?

You should be able to use the ExpiFectamine293 Reagent for transfection of Freestyle 293-F cells grown in FreeStyle 293 Expression Medium; however, the enhancers will provide only a little boost in expression as they are designed to work with higher density cultures that FreeStyle 293 Expression Medium cannot support.

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

Are there any recommendations for preventing or dispersing cell clumps in a suspension culture?

When cells meant to be grown in suspension are grown in static culture, they may form clumps. These clumps will severely limit transfection efficiency and protein expression. It is suggested that FreeStyle 293 cells in FreeStyle media and CHO-S cells in CD-CHO or CHO-SFM are grown in agitated suspension to reduce the appearance of clumps. However, if clumps do form, you can try the following protocol to select for cells that don't form clumps:

- Transfer cells into an appropriate size centrifuge tube that will hold the entire cell suspension.
- Allow cells to sit undisturbed for about 5 minutes. The time can vary depending on the specific cell line. Larger cell clumps will settle to the bottom of the tube.
- Collect cells from the upper portion of the tube to passage into a new flask.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Is it necessary to grow FreeStyle 293-F cells in 8% CO2 when using the FreeStyle 293 Expression Medium? If 8% CO2 cannot be arranged, can I supplement the media with an additional reagent?

Our recommendation of 8% CO2 for the optimal growth of Freestyle 293 cells in Freestyle 293 Expression Medium is based on the bicarbonate content in the media and growth conditions of the cells observed by R&D. That said, you may use lower concentrations of CO2 if necessary (5% is common) to grow these cells. You may observe slightly slower growth kinetics or lower cell concentrations, but functionally the entire system will still work.

Does FreeStyle 293 Expression Medium contain any components that would interfere with concentrating secreted proteins from HEK 293 cells via ammonium sulfate precipitation?

Yes, the FreeStyle 293 Expression Medium contains Pluronic F-68 and lipids that would interfere with ammonium sulfate precipitation. Dialysis or tangential flow filtration can be used to remove lipids and Pluronic F-68 from the medium. Then you can concentrate the medium with ammonium sulfate.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.