商品编号


EG-42



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




100 μg

商品信息



Description

Sortase A is a bifunctional protein displaying both protein ligase/transpeptidase and cysteine protease activities. The ligase acitivity allows protein ligation generating fusion proteins or non-peptidic bioconjugates, such as unnatural amino acids, nucleic acids, fluorophores, chemical dyes and ligands (biotin) , antibiotics, sugar derivatives, etc. The protease activity of sortase A facilitates the cleavage of fusion proteins joined by LPXTGG motifs. The cleavage products serve as substrates for protein ligation. Consequently, sortase A mediates reactions reach an assay-specific equilibrium between ligation and cleavage activities.



Applications



in vitro protein ligation: ligate of LPETGG-tagged target protein to gycine modified oligonucleotide or GGK-fluorophores

Immobilization of proteins to solid surfaces

Cell-surface labelling of living organisms

Protein circularization

Posttranslational incorporation of novel functionality in proteins

Cleavage of fusion proteins joined by LPXTGG motifs.





Source

Purified from a recombinant
E. coli
strain carrying the Staphylococcus aureus sortase A amino acids 26 – 206.



Components



Sortase A
, 1 mg/ml in storage buffer: 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C.





Enzymatic activity

Sortase reactions can not proceed with 100 % efficiency, neither with respect to protein ligase, nor to protease activity. Rather an assay specific equilibrium between ligated and cleaved product is obtained. In consequence, sortase assays will always yield a mixture of cleaved and ligated proteins. Thus, further purification steps are needed to purify fusion protein.



Quality Control



The absence of endo-, exodeoxyribonucleases confirmed by appropriate quality tests.

Functionally tested for the capacity to join two proteins





Storage

-20°C.