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SCR508

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TAT-CRE Recombinase

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.

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Synonym(s):
Integrase, Recombinase Cre, cre recombinase
eCl@ss:
32160405
NACRES:
NA.75

biological source

Escherichia coli

Quality Level

recombinant

expressed in E. coli

assay

>70% (SDS-PAGE)

form

liquid

technique(s)

cell culture | stem cell: suitable

shipped in

dry ice

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This Item
C7988MAB3120CC130
vibrant-m

SCR508

TAT-CRE Recombinase

vibrant-m

C7988

Anti-Cre antibody, Mouse monoclonal

assay

>70% (SDS-PAGE)

assay

-

assay

-

assay

≥92% (SDS-PAGE)

biological source

Escherichia coli

biological source

mouse

biological source

mouse

biological source

Escherichia coli, human

form

liquid

form

buffered aqueous solution

form

-

form

liquid

shipped in

dry ice

shipped in

wet ice

shipped in

dry ice

shipped in

dry ice

Quality Level

100

Quality Level

200

Quality Level

100

Quality Level

100

General description

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.

Application

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.

Quality

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

Unit Definition

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.

Physical form

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

Storage and Stability

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 Class

10 - Combustible liquids

wgk_germany

WGK 1


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Raktima Raychowdhury et al.
ImmunoHorizons, 5(10), 818-829 (2021-10-21)
In this study, we report that the TLR4 ligand, LPS, and TLR3 ligand polyinosinic:polycytidylic acid failed to activate IRF3 or STAT1 in bone marrow-derived macrophages (BMMs) isolated from two independently generated lines of Rosa26-integrated Cas9-expressing C57BL/6J (B6) mice. RNA-sequencing analysis
Georgia Wilke et al.
Cell host & microbe, 26(1), 123-134 (2019-06-25)
Despite being a frequent cause of severe diarrheal disease in infants and an opportunistic infection in immunocompromised patients, Cryptosporidium research has lagged due to a lack of facile experimental methods. Here, we describe a platform for complete life cycle development and
Site-specific recombination in human embryonic stem cells induced by cell-permeant Cre recombinase.
Nolden, Lars, et al.
Nature Methods, 3, 461-467 (2006)
Mian Zhang et al.
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 34(4), 726-738 (2018-11-30)
Traumatic joint injuries produce osteoarthritic cartilage manifesting accelerated chondrocyte terminal differentiation and matrix degradation via unknown cellular and molecular mechanisms. Here we report the ability of biomechanical stress to increase expression of the calcium-sensing receptor (CaSR), a pivotal driver of
Stem cell engineering using transducible Cre recombinase.
Nolden, Lars, et al.
Methods in Molecular Medicine, 140, 17-32 (2007)

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