Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
PRO-320

Background :

Recombinant Listeriolysin O (LLO) is a single polypeptide protein secreted by the Gram-positive bacterium Listeria monocytogenes. LLO belongs to the group of cholesterol-binding sulfhydrylactivated toxins, the lytic activity of which is enhanced by reducing agents and is suppressed by exposure to oxygen or cholesterol. LLO hemolytic activity is maximum at pH 5.5 and rapidly decreases with the increase of the pH. Listeriolysin O is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered clear solution.

Formulation:

The protein (0.2mg/ml) contains 50mM NaH2PO4, 1mM EDTA, 2.7mM KCl, 1mM DTT, 5% glycerol and 0.5M NaCl.

Stability:

Recombinant Listeriolysin O although stable at room temperature for 1 week, should be stored desiccated below -18 C.
Please avoid freeze-thaw cycles.

Purity:

Greater than 90.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:

Recombinant LLO has a hemolytic activity 1.25x106 HU/mg. 2mM DTT is used to reactivate the toxin.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of LLO.

Applications:

a. Cytosolic delivery of molecules, peptides, oligonucleodides and plasmid DNA.
b. Production of specific monoclonal antibodies.
c. Detection of anti-listeriolysin O antibodies.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:hly
Synonyms:hlyA, lisA
OrderedLocusNames:GN:lmo0202#ORF">lmo0202

Protein synonyms/aliases:

Listeriolysin O precursor (Thiol-activated cytolysin)
    (LLO).

Protein Family:

Belongs to the thiol-activated cytolysin family.

Protein Domains:

Domains:
•IPR001869   Thiol-activated cytolysin

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Phosphorylation, ubiquitination and degradation of listeriolysin O in mammalian cells: role of the PEST-like sequence.
Cell Microbiol 2006 Feb;8(2):353-64
2. Listeria monocytogenes listeriolysin O and phosphatidylinositol-specific phospholipase C affect adherence to epithelial cells.
Can J Microbiol 2005 Sep;51(9):745-51
3. Detection of Listeria monocytogenes and the toxin listeriolysin O in food.
J Microbiol Methods 2006 Feb;64(2):141-70
4. Multiple synergizing factors contribute to the strength of the CD8+ T cell response against listeriolysin O.
Int Immunol 2006 Jan;18(1):89-100
5. Listeriolysin O, a cytolysin derived from Listeria monocytogenes, inhibits generation of ovalbumin-specific Th2 immune response by skewing maturation of antigen-specific T cells into Th1 cells.
Clin Exp Immunol 2005 Nov;142(2):268-74
6. Fusion to Listeriolysin O and delivery by Listeria monocytogenes enhances the immunogenicity of HER-2/neu and reveals subdominant epitopes in the FVB/N mouse.
J Immunol 2005 Sep 15;175(6):3663-73

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中華民國95年06月06日更新