Flp-In™-3T3 Cell Line

Flp-In™ Cell Lines are designed for rapid generation of stable cell lines that express a protein of interest from aRead more

Catalog Number	Quantity
R76107 also known as R761-07	1 mL

Catalog number R76107

also known as R761-07

Price (JPY)

208,200

Quantity:

Flp-In™ Cell Lines are designed for rapid generation of stable cell lines that express a protein of interest from a Flp-In™ expression vector. These cells contain a single stably integrated FRT site at a transcriptionally active genomic locus. Targeted integration of a Flp-In™ expression vector ensures high-level expression of your gene of interest. There are six Flp-In™ Cell Lines available for the generation of isogenic stable cell lines and a Flp-In™ T-REx™ Cell Line for the generation of tetracycline-regulated stable cell lines. Flp-In™-CV-1, Flp-In™-293, Flp-In™-BHK, Flp-In™-Jurkat, and Flp-In™-3T3 Cell Lines were created by transfecting the parent cell lines with pFRT/lacZeo and selecting for stable Zeocin™- resistant clones. The Flp-In™-CHO Cell Line was created by transfecting CHO cells with pFRT/lacZeo2 and selecting for Zeocin™-resistant clones. The Flp-In™ T-REx™-293 Cell Line contains pFRT/lacZeo and pcDNA™6/TR (from the T-REx™ System) stably integrated. Co-transfection of the Flp-In™ Cell Lines with a Flp-In™ expression vector and the Flp recombinase vector, pOG44, results in targeted integration of the expression vector to the same locus in every cell, ensuring homogeneous levels of gene expression.

Choosing your Flp-In™ Vector/Cell Line Combination
The Flp-In™-CV-1, Flp-In™-293, Flp-In™-CHO, and Flp-In™-Jurkat Cell Lines (Figure 1) work well with Flp-In™ vectors that express a gene from the CMV promoter (pcDNA™5/FRT, pcDNA5/FRT/V5-His-TOPO™, and pSecTag/FRT/V5-His-TOPO™). Flp-In™-BHK and Flp-In™-3T3 cells tend to down regulate the CMV promoter. Therefore, it is recommended that the Flp-In™ vectors containing the EF-1α promoter (pEF5/FRT/V5-DEST™ and pEF5/FRT/V5-D-TOPO™) be used with these cell lines.

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

Specifications

Cell LineFlp-In™-3T3

Quantity1 mL

SpeciesMouse

Product LineFlp-In

Protein TagpSec

Unit SizeEach

Contents & Storage

3 x 10⁶cells 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.

Frequently asked questions (FAQs)

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.

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

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.

Citations & References (1)

Citations & References

Abstract

Individual overexpression of five subunits of human translation initiation factor eIF3 promotes malignant transformation of immortal fibroblast cells.

Authors:Zhang L, Pan X, Hershey JW,

Journal:J Biol Chem

PubMed ID:17170115

Transcriptional and post-transcriptional regulatory mechanisms are commonly accepted paradigms of tumorigenesis. The view is emerging that deregulation of translation contributes importantly to cancer development, a role not generally appreciated before. Eukaryotic initiation factor eIF3 contains at least thirteen non-identical subunits, named from eIF3a to eIF3m, and plays an essential role ... More