Gibco™

Expi293™ Inducible GnTI- Expression System Kit

Catalog number:  A39252

 Related applications:

Mammalian Cell Culture

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Description

The Expi293 Inducible GnTI- Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian 293 cells. The system kit is centered on engineered Expi293F cells that do not have N-acetylglucosaminyltransferase I (GnTI) activity, therefore lacking complex N-glycans, and stably express the tetracycline repressor (TetR) protein, thus enabling tetracycline-regulated protein expression. The Expi293F Inducible GnTI- cell line was established by the selective knockout of the GnTI gene and the stable integration of the regulatory plasmid pcDNA6/TR, which encodes the TetR gene under the control of the human CMV promoter, in parental Expi293F cells. This cell line is a powerful tool for tetracycline-inducible high-yield expression of homogeneously glycosylated recombinant proteins.

Expi293 Inducible GnTI- Expression System Kit features include:
• Homogeneous N-glycosylation of expressed proteins
• Tetracycline-inducible protein expression with extremely low basal expression levels
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• pcDNA5/TO expression vector for tetracycline-regulated expression of the gene of interest
• Suspension, high-density culture of Expi293F Inducible GnTI- cells in Expi293 Expression Medium
• High-efficiency transient transfection using ExpiFectamine 293 reagent in combination with specialized transfection enhancers

The components included in the Expi293 Inducible GnTI- Expression System Kit are: one vial of frozen Expi293F Inducible GnTI- cells, 1 L of Expi293 Expression Medium, one ExpiFectamine 293 transfection kit sufficient to transfect 1 L of culture, 100 mL Opti-Plex Complexation Buffer, one PNGase F Glycan Cleavage kit, one pcDNA5/TO Mammalian Expression Vector kit, 500 mg tetracycline hydrochloride, and the pRABBIT IgG IRES-EmGFP positive control vector. All components of the Expi293 Inducible GnTI- Expression System are also available for individual purchase.

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

Specifications

Cell Line: Expi293F™
Delivery Method: Transfection
Key Function: Protein Production
Product Line: Expi293™
Product Size: 1 kit
Promoter: CMV⁄TO
Shipping Condition: Approved for shipment at Room Temperature or on Wet or Dry Ice

Contents & storage

• Expi293F Inducible GnTI- Cells (1x107 cells/vial), store in vapor-phase liquid nitrogen
• 1 L Expi293 Expression Medium, store at 2–8°C (protect from light)
• ExpiFectamine 293 Transfection Kit for 1 L of culture, store at 2–8°C
• PNGase F Glycan Cleavage Kit, store at -20C
• 100 mL Opti-Plex Complexation Buffer, store at 2–8°C (protect from light)
• 20 µg pcDNA5/TO Mammalian Expression Vector, store at 2–8°C or long term storage at -20°C.
• 500 mg Tetracycline Hydrochloride, store at -20°C
• pRABBIT IgG IRES-EmGFP Positive Control Vector, store at 2–8°C or long term storage at -20°C

Documents

Manuals & protocols