Expi293F™ GnTI- Cells

La línea de células Expi293F GnTI- se deriva de las células Expi293F diseñadas que no tienen actividad de N-acetilglucosaminiltransferasa IMás información

Número de catálogo	Cantidad
A39240	1 vial

Número de catálogo A39240

Precio (CLP)

Cantidad:

1 vial

La línea de células Expi293F GnTI- se deriva de las células Expi293F diseñadas que no tienen actividad de N-acetilglucosaminiltransferasa I (GnTI), y por lo tanto carecen de N-glicanos complejos. Es una potente herramienta para la expresión de alto rendimiento de proteínas recombinantes homogéneamente glicosiladas.

Características de las células Expi293F GnTI-:
• Suspensión, cultivo de alta densidad en medio de expresión Expi293
• N-glicosilación homogénea de proteínas expresadas
• Rendimiento proteínas equivalente al de las células parentales Expi293F: hasta 1 g/l
• Rendimiento de proteínas significativamente más altos que el de otras líneas de células alternativas HEK293 GnTI-

Las células congeladas se suministran en un vial que contiene 1 ml de células a 1 x 10⁷ células viables/ml en el medio de expresión Expi293 y dimetilsulfóxido (DMSO) al 10 %. Las células se deben descongelar directamente en el medio de expresión Expi293.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Línea de productosExpi293

Cantidad1 vial

Condiciones de envíoHielo seco

Unit SizeEach

Contenido y almacenamiento

1 vial de células Expi293F GnTI- (1 x 10⁷ células/vial), almacenar en nitrógeno líquido en fase de vapor

Preguntas frecuentes

Can I leave my Expi293F culture expressing for more than 7 days?

The growth and expression characteristics of Expi293F cells are such that, by 7 days post-transfection, the culture medium should be close to being spent and maximal protein expression should have already been achieved. Continued incubation will result in a large decrease in cell culture viability.

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

If I let my Expi293F cells grow to a higher density than recommended, will they be okay?

While the Expi293 Expression Medium can support much higher cell densities, we do not recommend growing your Expi293F cultures beyond 5-6 x 10e6 cells/mL, as subsequent transfection and protein expression efficiencies may be reduced. At higher densities, there is also the increased possibility of reaching the point of reduced culture viability. If your seed culture does exceed 5-6 x 10e6 cells/mL, passage them once or twice as detailed in the manual, monitoring them for viability and growth rate. Perform a test transfection using the Protein Expression Control IgG or an expression construct of known yield to determine if cell expression performance has been impacted.

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

What method do you recommend for quantifying rabbit IgG expression from pRABBIT IgG IRES-EmGFP Positive Control Vector, Cat. No. A39243?

The formation of intact IgG molecules may be quantified using a sandwich ELISA designed to capture and detect rabbit IgG. Besides the rabbit IgG positive control vector, reagents, and consumables that are included in the kit, you will also need purified rabbit IgG to be used as a standard, F(ab')2-Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Cat. No. A10547), Protein A Coated Plates, Clear, 96-Well (Cat. No. 15130), TMB Substrate Kit (Cat. No. 34021), SuperBlock (TBS) Blocking Buffer (Cat. No. 37535), and PBS or TBS buffer for washes. There is an example procedure in our Protein A Coated Plates manual (https://tools.thermofisher.com/content/sfs/manuals/MAN0011310_Thermo Scientific_ProteinA_G_AG_Coat_96Well_UG.pdf). Please note that our R&D scientists determine titer values from crude cell culture supernatants using a Pall Life Sciences FortéBio Octet instrument equipped with a Protein A biosensor.

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

Do you offer a GFP-expressing mammalian expression vector that I can use as a control to monitor my transfection and expression with the Expi systems?

We offer pRABBIT IgG IRES-EmGFP Positive Control Vector, Cat. No. A39243, which you can use to monitor your transfection and expression.

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

How do I culture Expi293F GnTI-, Expi293F Inducible, or Expi293F Inducible GnTI- cells?

Expi293F GnTI-, Expi293F Inducible, and Expi293F Inducible GnTI- cell lines were derived from the parental Expi293F Cells (Cat. No. A14527) and therefore, share similar morphology and growth characteristics. They are cultured in Expi293 Expression Medium and transfected using ExpiFectamine 293 Transfection Kit following our standard Expi293 protocol.

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

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