TAT-Cre Recombinase

Description
Catalogue Number	SCR508
Description	TAT-CRE Recombinase
Alternate Names	Integrase Recombinase Cre
Background Information	Cre Recombinase is an enzyme from bacteriophage P1 that catalyzes the site-specific recombination between two DNA recognition sites termed loxP sites. The LoxP recognition site consists of two 13 bp inverted repeats flanking a 8 bp spacer region. Because the Cre gene and loxP sites are not native to the genomes of most species, LoxP sites can be engineered and introduced into target cells and thus used as a means to precisely control the expression of genes in vitro (i.e. cultured cells) and in vivo (i.e. animal models). • If LoxP sites are located on different chromosomes, Cre recombinase will mediate a chromosomal translocation. • If LoxP sites are oriented in the opposite direction, Cre recombinase will mediate an inversion of the floxed segment. • If LoxP sites are oriented in the same direction, Cre recombinase will mediate a deletion of the floxed segment. In this way, placement of the LoxP sites allows genes to be activated, repressed or exchanged for other genes. EMD Millipore’s TAT-CRE Recombinase is a recombinant cell-permeant fusion protein consisting of a basic protein translocation peptide derived from HIV-TAT (TAT), a nuclear localization sequence (NLS), the Cre protein and an N-terminal histidine tag (H6) for efficient purification of the protein from E. coli. EMD Millipore’s TAT-CRE Recombinase has been shown to effectively excise STEMCCA viral transgenes from both Human and Mouse IPS cells.

Description

Catalogue Number

SCR508

Description

TAT-CRE Recombinase

Alternate Names

Integrase
Recombinase Cre

Background Information

Cre Recombinase is an enzyme from bacteriophage P1 that catalyzes the site-specific recombination between two DNA recognition sites termed loxP sites. The LoxP recognition site consists of two 13 bp inverted repeats flanking a 8 bp spacer region. Because the Cre gene and loxP sites are not native to the genomes of most species, LoxP sites can be engineered and introduced into target cells and thus used as a means to precisely control the expression of genes in vitro (i.e. cultured cells) and in vivo (i.e. animal models).

• If LoxP sites are located on different chromosomes, Cre recombinase will mediate a chromosomal translocation.
• If LoxP sites are oriented in the opposite direction, Cre recombinase will mediate an inversion of the floxed segment.
• If LoxP sites are oriented in the same direction, Cre recombinase will mediate a deletion of the floxed segment.

In this way, placement of the LoxP sites allows genes to be activated, repressed or exchanged for other genes.

EMD Millipore’s TAT-CRE Recombinase is a recombinant cell-permeant fusion protein consisting of a basic protein translocation peptide derived from HIV-TAT (TAT), a nuclear localization sequence (NLS), the Cre protein and an N-terminal histidine tag (H6) for efficient purification of the protein from E. coli.

EMD Millipore’s TAT-CRE Recombinase has been shown to effectively excise STEMCCA viral transgenes from both Human and Mouse IPS cells.

Product Information
Presentation	Recombinant protein is supplied in buffer containing 50% glycerol (v/v) 500 mM NaCl and 20 mM HEPES at pH 7.4
Activity	A standard of 100 Units is defined as the amount of TAT-CRE (ug) in 1.0 mL of tissue culture medium that is required to induce 50% GFP expression in a HEK293T loxp reporter cell line assay.

Product Information

Presentation

Recombinant protein is supplied in buffer containing 50% glycerol (v/v) 500 mM NaCl and 20 mM HEPES at pH 7.4

Activity

A standard of 100 Units is defined as the amount of TAT-CRE (ug) in 1.0 mL of tissue culture medium that is required to induce 50% GFP expression in a HEK293T loxp reporter cell line assay.

Applications
Application	TAT-CRE Recombinase is a recombinant cell-permeant fusion cre-recombinase protein consisting of TAT sequence, a nuclear localization sequence (NLS) and it is known to catalyze the site specific recombination event between two loxP DNA sites.
Key Applications	Cell Culture Stem Cell Culture
Application Notes	EMD Millipore has developed a cell-permeant TAT-CRE Recombinase fusion protein which can be directly delivered to mammalian cells and results in high recombination efficiencies (75 – 100% in mouse and ~ 60% in human cells). TAT-CRE readily translocates to mammalian cells and can catalyze highly efficient recombination. The dose and timing of Cre exposure can be precisely controlled thus allowing for the careful titration of Cre activity.

Applications

Application

TAT-CRE Recombinase is a recombinant cell-permeant fusion cre-recombinase protein consisting of TAT sequence, a nuclear localization sequence (NLS) and it is known to catalyze the site specific recombination event between two loxP DNA sites.

Key Applications

Cell Culture
Stem Cell Culture

Application Notes

EMD Millipore has developed a cell-permeant TAT-CRE Recombinase fusion protein which can be directly delivered to mammalian cells and results in high recombination efficiencies (75 – 100% in mouse and ~ 60% in human cells). TAT-CRE readily translocates to mammalian cells and can catalyze highly efficient recombination. The dose and timing of Cre exposure can be precisely controlled thus allowing for the careful titration of Cre activity.

Biological Information
Host	E. coli

Biological Information

Host

E. coli

Physicochemical Information
Activity	A standard of 100 Units is defined as the amount of TAT-CRE (ug) in 1.0 mL of tissue culture medium that is required to induce 50% GFP expression in a HEK293T loxp reporter cell line assay.

Physicochemical Information

Activity

A standard of 100 Units is defined as the amount of TAT-CRE (ug) in 1.0 mL of tissue culture medium that is required to induce 50% GFP expression in a HEK293T loxp reporter cell line assay.

Product Usage Statements
Quality Assurance	Each lot of TAT-CRE Recombinase protein is rigorously quality control tested for the following parameters: • Purity: single band around 41 kDa with greater than 70% protein purity on an SDS-PAGE gel • Functional activity: mediates recombination of LoxP-modified alleles in a HEK293T- Cre reporter cell line • Endotoxin levels: less than 1 EU/ug protein • Mycoplasma negative
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Each lot of TAT-CRE Recombinase protein is rigorously quality control tested for the following parameters:
• Purity: single band around 41 kDa with greater than 70% protein purity on an SDS-PAGE gel
• Functional activity: mediates recombination of LoxP-modified alleles in a HEK293T- Cre reporter cell line
• Endotoxin levels: less than 1 EU/ug protein
• Mycoplasma negative

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 3 months from date of receipt when stored at -20°C or -80°C. Upon first thaw, centrifuge the vial and gently mix the solution. Aliquot into smaller working volumes and freeze at -20°C or -80°C. Before use, dilute TAT-CRE to the appropriate concentration with culture medium and filter through a 0.2um low protein binding syringe filter (Millipore Cat. No. SLGV 033RS or SLGV013 SL).

Storage and Shipping Information

Storage Conditions

Stable for 3 months from date of receipt when stored at -20°C or -80°C. Upon first thaw, centrifuge the vial and gently mix the solution. Aliquot into smaller working volumes and freeze at -20°C or -80°C. Before use, dilute TAT-CRE to the appropriate concentration with culture medium and filter through a 0.2um low protein binding syringe filter (Millipore Cat. No. SLGV 033RS or SLGV013 SL).

Packaging Information
Material Size	1500 units

Packaging Information

Material Size

1500 units

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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