Gibco™

EXPI293™ Induzierbares Expressionssystemkit

Katalognummer	Menge
A39251	1 Kit

Katalognummer A39251

Preis (EUR)

3.310,00

Menge:

1 Kit

Preis (EUR)

3.310,00

Das Expi293 induzierbare Expressionssystemkit ist Teil der Expi293 Plattform zur schnellen, ergiebigen, transienten Proteinexpression aus Säugetier-293-Zellen. Das Systemkit ist auf speziell entwickelte Expi293F Zellen ausgelegt, die das Tetracyclin-Repressor-(TetR)-Protein stabil exprimieren und so eine Tetracyclin-regulierte Proteinexpression ermöglichen Die Expi293F induzierbare Zelllinie wurde durch die stabile Integration des regulatorischen Plasmids pcDNA6/TR etabliert, das das TetR-Gen unter der Kontrolle des menschlichen CMV-Promotors kodiert Diese Zelllinie ist ein leistungsstarkes Werkzeug zur ertragreichen Tetracyclin-induzierbaren Expression rekombinanter Proteine.

Merkmale des Expi293 induzierbaren Expressionssystemkits:
• Tetracyclin-induzierbare Proteinexpression mit extrem niedrigen Basalexpressionswerten
• Ähnliche Proteinausbeute wie bei elterlichen Expi293F Zellen — bis zu 1 g/l
• pcDNA5/TO-Expressionsvektor zur Tetracyclin-regulierten Expression des Gens von Interesse
• Suspension, hochdichte Kultur von Expi293F induzierbaren Zellen im Expi293 Expressionsmedium
• Hocheffiziente transiente Transfektion mit ExpiFectamine 293 Reagenz in Kombination mit speziellen Transfektionsverstärkern

Inhalt des Expi293 induzierbaren Expressionssystemkits: ein Fläschchen mit gefrorenen Expi293F induzierbaren Zellen, 1 l Expi293 Expressionsmedium, ein ExpiFectamine 293 Transfektionskit, ausreichend zur Transfektion von 1 l Kultur, 100 ml Opti-Plex Komplexationspuffer, ein pcDNA5/TO-Säugetier-Expressionsvektorkit, 500 mg Tetracyclin-Hydrochlorid und pRABBIT IgG IRES-EmGFP positiver Kontrollvektor. Alle Komponenten des Expi293 induzierbaren Expressionssystems sind auch separat erhältlich.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

ZelllinieExpi293F™

LiefertypTransfektion

Zur Verwendung mit (Anwendung)Proteinexpression

ProduktlinieExpi293

Menge1 Kit

VersandbedingungFreigegeben für den Versand ohne Kühlung oder auf nassem Eis oder Trockeneis

AusgangsmaterialDNA

PromoterCMV/TO

TypInduzierbares Expressionssystemkit

Unit SizeEach

Inhalt und Lagerung

• Expi293F induzierbare Zellen (1 x 10⁷ Zellen/Fläschchen), Lagerung in Stickstoffgas
• 1 l Expi293 Expressionsmedium, Lagerung bei 2 – 8 °C (vor Licht schützen)
• ExpiFectamine 293 Transfektionskit für 1 l Kultur, Lagerung bei 2 – 8 °C
• 100 ml Opti-Plex Komplexierungspuffer, Lagerung bei 2 – 8 °C (vor Licht schützen)
• 20 µg pcDNA5/TO-Säugetier-Expressionsvektor, Lagerung bei 2 – 8 °C oder längere Lagerung bei -20 °C.
• pRABBIT IgG IRES-EmGFP positiver Kontrollvektor, Lagerung bei 2 – 8 °C oder längere Lagerung bei -20 °C.
• 500 mg Tetracyclin-Hydrochlorid, Lagerung bei -20 °C

Häufig gestellte Fragen (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.

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.

Can I use doxycycline instead of tetracycline as an inducer in the Expi293 Inducible Expression System?

Doxycycline may be used as an alternative inducing agent in the Expi293 Inducible System. It is similar to tetracycline in its mechanism of action. Doxycycline has been shown to have a longer half-life than tetracycline (48 hr vs. 24 hr, respectively). If the use of doxycycline is desired with the Expi293 Inducible System, we recommend performing a dose-response curve to determine the optimal concentration for induction.

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

View all EXPI293™ Induzierbares Expressionssystemkit FAQs