商品编号


EG-1002



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Genome Engineering




1 mg

商品信息



Description


FLP recombinase catalyzes the site-specific recombination of DNA between FRT sites. It has been widely used for generating conditional knockout mice, mediated by FLP/FRT system.

Recombinant cell-permeant FLP recombinase Tat-Flp was purified from an HEK293 cell line overexpressing FLP Recombinase from yeast
Saccharomyces cerevisiae
with additional N-terminal 6XHis tag, a Tat peptide (GRKKRRQRRRPPAGTSVSL) and an NLS sequence (PKKKRKV). In addition, this "
Thermo
stable" mutant FLP (P2S L33S Y108N S294P) has ~ 4-10 fold higher activity than wild-type FLP at 37°C.




Applications

ex vivo
FRT recombination.



QC


Addition of 100 units of Tat-Flp to FRT-RFP-Stop-FRT-GFP 293T reporter cells resulted in 60~90% recombination efficiency.


Figure 1
: Tat-Flp transduction efficiency is concentration dependent.

1
: no Tat-FLP;
2~8
: adding 5, 10, 25, 50, 75, 100 and 200 units of Tat-FLP to HEK 293T cells transfected with FRT-RFP-Stop-FRT-GFP plasmid DNA. Pictures were taken 4 days after transduction with various amount of Tat-FLP




Unit Definition

A standard of 100 Units is defined as the amount of TAT-FLP in 1.0 mL of tissue culture medium that is required to induce 50%?GFP expression in a HEK 293T reporter cell line assay.



Source

Sf9 cells



Storage

-20 ~ -80 °C.