Gibco™

Expi293F™ Inducible GnTI- Cells

La lignée cellulaire GnTI inductible Expi293F est dérivée de cellules Expi293F conçues qui n’ont pas d’activité N-acétylglucosaminyltransférase I (GnTI), etAfficher plus

Référence	Quantité
A39242	1 flacon

Référence A39242

Prix (EUR)

3 085,00

Quantité:

1 flacon

Prix (EUR)

3 085,00

La lignée cellulaire GnTI inductible Expi293F est dérivée de cellules Expi293F conçues qui n’ont pas d’activité N-acétylglucosaminyltransférase I (GnTI), et qui manquent donc de N-glycanes complexes. Elles expriment de façon stable la protéine répresseur de tétracycline (TetR), permettant ainsi l’expression protéique régulée par la tétracycline. La lignée cellulaire a été établie par la neutralisation sélective du gène GnTl et l’intégration stable du plasmide de régulation pcDNA6/TR, qui code le gène TetR sous le contrôle du promoteur humain CMV. C’est un outil puissant pour l’expression à haut rendement inductible par la tétracycline de protéines recombinantes glycosylées homogènes.

Caractéristiques des cellules GnTI inductibles Expi293F :
• Culture en suspension, à haute densité dans le milieu d’expression Expi293
• N-glycosylation homogène des protéines exprimées
• Expression protéique inductible par la tétracycline avec des niveaux d’expression basale extrêmement faibles
• Rendements protéiques équivalents à la cellule parentale Expi293F : jusqu’à 1 g / l

Pour l’expression inductible de votre protéine d’intérêt, le vecteur d’expression de pcDNA5/TO de mammifères ou un vecteur inductible équivalent est requis pour l’expression génétique régulée par la tétracycline.

Les cellules congelées sont fournies dans un flacon contenant 1 ml de cellules à 1 x 10⁷ cellules viables / ml dans un milieu d’expression EXPI293 et 10 % de DMSO. Les cellules doivent être décongelées directement dans le milieu d’expression Expi293.

Usage exclusivement réservé à la recherche. Ne pas utiliser pour des procédures de diagnostic.

Spécifications

Gamme de produitsExpi293

Quantité1 flacon

Conditions d’expéditionGlace carbonique

Unit SizeEach

Contenu et stockage

1 flacon de cellules GnTI inductible Expi293F (1 x 10⁷ cellules / flacon), stocker dans de l’azote liquide en phase vapeur

Foire aux questions (FAQ)

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 is the difference between the Expi293F Inducible Cell Line and the T-REx -293 Cell Line?

The Expi293F Inducible Cell Line was created using the same technology as the T-REx -293 Cell Line. Both cell lines have been created by stable transfection with the pcDNA 6/TR vector; thus, they ubiquitously express the tetracycline repressor (TetR) protein. The parental cell line of Expi293F Inducible cells is Expi293F Cells (Cat. No. A14527). The key differences between these 2 cell lines are:
- Expi293F Inducible cells are grown in suspension and are adapted to high-density culture (> 15x10E6 cells/mL).
- Expi293F Inducible cells grow in Expi293 Expression System and perform equivalently to parental Expi293F cells, thus making them ideal for high-yield, inducible protein expression.

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.

View all Expi293F™ Inducible GnTI- Cells FAQs