FreeStyle™ 293 Expression System
Product Image
Citas y referencias (5)
Invitrogen™

FreeStyle™ 293 Expression System

El sistema de expresión FreeStyle™ 293 es un completo sistema de cultivo celular en suspensión para la generación de grandesMás información
Have Questions?
Número de catálogoCantidad
K9000011 Kit
Número de catálogo K900001
Precio (MXN)
-
Cantidad:
1 Kit
El sistema de expresión FreeStyle™ 293 es un completo sistema de cultivo celular en suspensión para la generación de grandes cantidades de proteínas recombinantes de mamíferos. Libere a su laboratorio del proceso laborioso y que requiere tiempo de sembrar, alimentar, pasar y mantener múltiples matraces de células adherentes (Figura 1). El sistema de expresión FreeStyle™ 293 combina el medio de expresión GIBCO™ FreeStyle™ 293 con 293fectin™, una formulación de lípidos-catiónicos diseñada para transfectar células FreeStyle™ 293-F en suspensión con alta eficacia. Las células 293-F Freestyle™ y el 293fectin™ están disponibles como parte del sistema de expresión o por separado

El medio de expresión FreeStyle™ 293 es un medio químico definido sin proteínas desarrollado específicamente por su capacidad para apoyar el crecimiento y la transfección de células 293-F en condiciones de cultivo de suspensión (Figura 2). Se trata de un medio completo, listo para su uso y sin materiales de origen animal que se ha suplementado con el suplemento GlutaMAX™-I. El medio de expresión Freestyle™ 293 permite ahorrar tiempo y costes significativos asociados a la adaptación de cultivos celulares a condiciones sin suero.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Línea de célulasCélulas FreeStyle™ 293F
ClasificaciónSin suero
Concentración1 x 10^7 células/ml
Tipo de cultivoCultivos celulares en suspensión
Tipo de entregaTransfección
Mecanismo de expresiónExpresión basada en células
Sistema de expresiónMamífero
Para utilizar con (aplicación)Expresión de proteínas
de control principalProducción de proteínas
Línea de productosFreestyle
Tipo de productoSistema de expresión
Cantidad1 Kit
Tipo de célulaCélulas derivadas 293
Unit SizeEach
Contenido y almacenamiento
El sistema de expresión FreeStyle™ 293 contiene:
  • Medio de expresión 1 l GIBCO™ FreeStyle™ 293
  • 1 x 107 células Freestyle™ 293-F (congeladas)
  • 1 ml de reactivo para transfección 293fectin™
  • 100 ml de Opti-MEM™ I para la formación de complejos
  • 25 µg de pCMV-SPORT β-gal
Consulte el Manual del sistema Freestyle™ 293 para obtener información sobre el almacenamiento y la manipulación. Almacene las células 293-F FreeStyle™ en nitrógeno líquido. Almacene los reactivos para transfección 293fectin™ a 4 °C. Almacene los medios de expresión GIBCO™ Freestyle™ 293 y Opti-MEM™ I a 2-8 °C en la oscuridad. Almacene PCMV SPORT β-gal a -20 °C. Se garantiza la estabilidad de los reactivos durante seis meses cuando se almacenan adecuadamente.

Preguntas frecuentes

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.

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

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).

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

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).

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

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.

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

Citations & References (5)

Citations & References
Abstract
Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10.
Authors:Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM, Sekiguchi K,
Journal:J Biol Chem
PubMed ID:14701821
'The adhesive interactions of cells with laminins are mediated by integrins and non-integrin-type receptors such as alpha-dystroglycan and syndecans. Laminins bind to these receptors at the C-terminal globular domain of their alpha chains, but the regions recognized by these receptors have not been mapped precisely. In this study, we sought ... More
Biological activity of follistatin isoforms and follistatin-like-3 is dependent on differential cell surface binding and specificity for activin, myostatin, and bone morphogenetic proteins.
Authors:Sidis Y, Mukherjee A, Keutmann H, Delbaere A, Sadatsuki M, Schneyer A,
Journal:Endocrinology
PubMed ID:16627583
Follistatin (FST) and FST-like-3 (FSTL3) are activin-binding and neutralization proteins that also bind myostatin. Three FST isoforms have been described that differ in tissue distribution and cell-surface binding activity, suggesting that the FST isoforms and FSTL3 may have some nonoverlapping biological actions. We produced recombinant FST isoforms and FSTL3 and ... More
Mutation of key residues of RPE65 abolishes its enzymatic role as isomerohydrolase in the visual cycle.
Authors:Redmond TM, Poliakov E, Yu S, Tsai JY, Lu Z, Gentleman S,
Journal:Proc Natl Acad Sci U S A
PubMed ID:16150724
RPE65 is essential for isomerization of vitamin A to the visual chromophore. Mutations in RPE65 cause early-onset blindness, and Rpe65-deficient mice lack 11-cis-retinal but overaccumulate alltrans-retinyl esters in the retinal pigment epithelium (RPE). RPE65 is proposed to be a substrate chaperone but may have an enzymatic role because it is ... More
Binding of anthrax toxin to its receptor is similar to alpha integrin-ligand interactions.
Authors:Bradley KA, Mogridge J, Jonah G, Rainey A, Batty S, Young JA,
Journal:J Biol Chem
PubMed ID:14507921
The secreted protein toxin produced by Bacillus anthracis contributes to virulence of this pathogen and can cause many of the symptoms seen during an anthrax infection, including shock and sudden death. The cell-binding component of anthrax toxin, protective antigen, mediates entry of the toxin into cells by first binding directly ... More
A novel secreted endonuclease from Culex quinquefasciatus salivary glands.
Authors:Calvo E, Ribeiro JM,
Journal:J Exp Biol
PubMed ID:16809456
Previous analysis of the salivary gland transcriptome of Culex quinquefasciatus showed the potential presence of an endonuclease with sequence similarities to shrimp, crab and two tsetse salivary proteins. Indeed, not only was the cloned cDNA shown to encode an active double-stranded endonuclease, but also the same activity was demonstrated to ... More