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Invitrogen™ Flp-In™-293 Cell Line

Catalog No. R75007
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R75007 1 mL
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Catalog No. R75007 Supplier Invitrogen™ Supplier No. R75007
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Includes

1 x 107 cells are supplied frozen in 1mL of 90% complete medium and 10% DMSO. Cells must be stored in liquid nitrogen.

Cells contain a single stably integrated FRT site at a transcriptionally active genomic locus

Flp-In 293 cell line is designed for rapid generation of stable cell lines that ensure high level expression of your protein of interest from a Flp-In expression vector. Targeted integration of a Flp-In expression vector ensures high-level expression of your gene of interest. Flp-In-293 cell line was created by transfecting the parent cell lines with pFRT/lacZeo and selecting for stable Zeocin™- resistant clones. Flp-In-293 cells work well with Flp-In vectors that express a gene from the CMV promoter (e.g., pcDNA™5/FRT, pcDNA5/FRT/V5-His-TOPO™, and pSecTag/FRT/V5-His-TOPO

Constitutive Expression, Mammalian Expression, Mammalian Expression Cell Lines, Protein Expression, Proteins, Expression, Isolation and Analysis, Targeted Integration

Specifications

Content And Storage 1 x 10E7 cells are supplied frozen in 1 ml of 90% complete medium and 10% DMSO. Cells must be stored in liquid nitrogen. Cells are guaranteed stable for 6 months when properly stored.
Cell Line Flp-In-293
Species Human
No. of Cells 1 mL
Product Line Flp-In
Quantity 1 mL
Is multiple integration of the Flp-In expression construct possible? How do you screen for multiple integrants, and how stable is the Flp-In expression cell line?

In theory, one can get multiple integrations of the Flp-In expression construct—an FRT-specific integration event and a random, second-site integration. However, random integration is a relatively uncommon event. Limiting the amount of DNA in the transfection will reduce the chance of second-site integration. We have transfected 293 cells (lacking the FRT site) with the pcDNA5/FRT vector and have identified one potential second-site integrant after screening over 200 clones. DNA integrations can be detected by Southern blot. A single integrant will display a single band; double: two; triple: three, etc. We have maintained a number of Flp-In expression cell lines for over four months and have not observed any loss of the Flp-In expression construct, whether hygromycin selection was maintained or not.

What kind of Flp-In T-REx cell lines do you offer?

We offer the Flp-In T-REx system that contains pFRT/lacZeo and pcDNA6/TR stably integrated into HEK 293 cells. This cell line has been functionally tested for its ability to regulate expression.

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