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Invitrogen™ FreeStyle™ 293 Expression System

Catalog No. K900001
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Description

Includes

The FreeStyle™ 293 Expression System contains: 1L GIBCO FreeStyle 293 Expression Medium, 1 x 107 FreeStyle 293-F Cells (frozen), 1mL 293fectin Transfection Reagent, 100mL Opti-MEM™ I for Complex Formation, 25μg pCMV-SPORT β-gal

Complete, suspension cell culture system for generating large amounts of mammalian recombinant protein

  • Combines GIBCO™ FreeStyle 293 Expression Medium with 293fectin™, a cationic-lipid formulation designed to transfect suspension FreeStyle 293-F cells at high efficiency
  • FreeStyle 293-F cells and 293fectin are available as part of the Expression System or separately
  • Chemically-defined, protein-free medium specifically developed for the ability to support the growth and transfection of 293-F cells under suspension type culture conditions
  • Complete, ready-to-use medium supplemented with animal-origin free GlutaMAX™-I Supplement
  • Saves time and costs associated with adaptation of cell cultures to serum-free conditions

Antibody & Cell Culture Production, Bioproduction, Cell Culture, Mammalian Expression, Mammalian Protein Production, Plasmid Transfection, Protein Expression, Proteins, Expression, Isolation & Analysis, Transfection, Transient Protein Production & High-Throughput Screening

Specifications

Cell Line FreeStyle™ 293-F Cells
Classification Serum-Free
Content And Storage The FreeStyle™ 293 Expression System contains:
  • 1L GIBCO™ FreeStyle™ 293 Expression Medium
  • 1 x 107 FreeStyle™ 293-F Cells (frozen)
  • 1 mL 293fectin™ Transfection Reagent
  • 100 mL Opti-MEM™ I for Complex Formation
  • 25 μg pCMV-SPORT β-gal
Please review the FreeStyle™ 293 System Manual for storage and handling information. Store FreeStyle™ 293-F Cells in liquid nitrogen. Store 293fectin™ Transfection Reagents at 4°C. Store GIBCO™ FreeStyle™ 293 Expression Medium and Opti-MEM™ I at 2-8°C in the dark. Store pCMV SPORT β-gal at -20°C. Reagents are guaranteed stable for six months when stored properly.
Delivery Type Transfection
Culture Type Suspension Cell Culture
Cell Type 293-derived Cells
Concentration 1 x 10^7 cells/mL
Expression Mechanism Cell-Based Expression
Expression System Mammalian
For Use With (Application) Protein Expression
Key Functions Protein Production
Product Line Freestyle
Product Type Expression System
Quantity 1 Kit
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Frequently Asked Questions (FAQs)

I am using your FreeStyle Max 293 Expression System and my cell viability is at most 87-90%. Is this normal? Do you have any tips to increase the cell viability? I am using 125 mL and 250 mL non-baffled, polycarbonate shaker flasks with vented caps. Will using baffled flasks help? Should I add Pluronic reagent to the medium?

Culture viability should be in the high 90% range for the seed stock that you are maintaining and growing for the transfections. After transfection, the viability will slowly decrease to the lower 90% to 80% range by about 3-6 days post-transfection and can drop more dramatically at 7 days or longer. This is when using our recommended protocol, which does not suggest any media changes or supplementation post-transfection.

A few parameters we recommend checking to assure high culture viability are: pH at ˜7.0, no visible clumping of the cells, CO2 levels at approximately 7-8% (this should be adjusted to achieve the appropriate pH), volume of culture at 40% or less of nominal flask capacity, and incubator temperature at 37 degrees C. If you have a shaker platform setup in your incubator, the platform may generate sufficient heat to increase the temperature of the incubator. We recommend using a thermometer attached to the shaker platform. Use of baffled flasks may help, if insufficient oxygenation of the cultures is suspected. We also recommend handling and storing the culture media properly, as light exposure can result in breakdown of certain media components. If you are using FreeStyle 293 Expression Medium, supplementing with Pluronic reagent is not necessary.
What is the main difference between the FreeStyle Max 293 Expression System and the FreeStyle 293 Expression System?

The two systems have the same components except for the transfection reagent and the medium supplied for formation of transfection complexes. The FreeStyle Max 293 Expression System contains FreeStyle Max Reagent and OptiPRO SFM (serum-free medium that is devoid of animal-origin components), whereas the FreeStyle 293 Expression System contains 293fectin Transfection Reagent and Opti-MEM I Reduced Serum Medium (serum-free medium but not animal-origin free).
Which vectors do you recommend for use with FreeStyle system?

Generally, any mammalian expression vector should be compatible with the FreeStyle expression system. Some of the more popular choices include pcDNA3.3-TOPO TA vector (Cat. No. K8300-01), pcDNA3.4-TOPO TA vector (Cat. no. A14697), pcDNA3.2 DEST vector (Cat. No. 12489018), pCEP vector (Cat. No. V044-50), or pcDNA4/HisMax vector (Cat. No. K864-20).
What is the difference between FreeStyle 293 Expression Medium and FreeStyle F17 Expression Medium?

FreeStyle F17 Expression Medium was designed from a different base formulation than FreeStyle 293 Expression Medium. FreeStyle F17 medium shows equal or better performance than the FreeStye 293 medium for certain cell lines and proteins. It is capable of supporting the growth of 293 and CHO cells and offers a wider variety and selection of transient cell culture media options.

For Research Use Only. Not for use in diagnostic procedures.