discontinued

AR10506PU-L Outer Membrane -A / ompA

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1 mg / €1,340.00

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Outer Membrane -A / ompA

AR10506PU-L

Product Description for Outer Membrane -A / ompA

Outer Membrane -A / ompA.
Presentation: Purified

Properties for Outer Membrane -A / ompA

Product Category Proteins & Growth Factors
Quantity 1 mg
Presentation Purified
Source E. coli
Species (Protein) E. coli
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Proteins (de)

Datasheet Extract

Purity >98.0% as determined by both RP-HPLC and SDS-PAGE analysis. 
Purification Method: Proprietary chromatographic techniques.
Background

The OmpA protein is one of the main outer-membrane proteins of a large array of Gram-negative bacteria such as A. salmonicida, Shigella dysenteriae and E. coli. OmpA's major physiological functions include maintenance of the structural integrity and morphology of the cells and porin activity, as well as a role in conjugation and bacteriophage binding. Achromogenic atypical Aeromonas salmonicida is the causative agent of goldfish ulcer disease. Virulence of this bacterium is associated with the production of a paracrystalline outer membrane A-layer protein. The species specific structural gene for the monomeric form of A-protein was cloned into a pET-3d plasmid in order to express and produce a recombinant form of the protein in E. coli BL21(DE3). The induced protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by ion exchange chromatography on Q-Sepharose to over 95% pure monomeric protein.
Recombinant A-Protein was compared by biochemical, immunological and molecular methods with the A-Protein isolated from atypical A. salmonicida bacterial cells by the glycine and the membrane extraction methods.

Storage Prior to reconstitution store at 2-8°C for one month or desiccated below -18°C.
Following reconstitution store undiluted at 2-8°C for one month
or (in aliquots) at -20°C for longer. 
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). 
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Description
Description:
The recombinant form was found to be undistinguishable from the wild type when examined by SDS-PAGE and gel filtration chromatography yielding a 48 kDa monomeric protein. The immunological similarity of the protein samples was demonstrated by employing polyclonal and monoclonal antibodies in ELISA and Western Blot techniques. All forms of A-Protein were found to activate the secretion of Tumour Necrosis Factor alpha from murine macrophage. For Ref see Maurice et al. (1999) Protein Expression and Purification 16, 396-404.
AA Sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Met-Asp-Val-Val-Ile-Ser.
Biological activity:
The interaction of bacterial and recombinant A-layer protein with murine macrophages was directed at determining the effect of A-protein on intracellular events that occur in primed macrophages. This was accomplished by measuring the cytotoxic product produced by peritoneal macrophages when exposed to A-protein coated latex beads. Thioglycolate elicited macrophages exhibited a low level of activation (18% cytotoxicity) that was significantly increased (48% cytotoxicity) in the presence of latex beads. Coating of the latex beads with each of the three A-protein products resulted in an increase of cytoxicity (mean +/- SEM) from 48% to 91%.
Format
Purity:
>98.0% as determined by both RP-HPLC and SDS-PAGE analysis. 
Purification Method: Proprietary chromatographic techniques.
State:
Sterile Filtered White lyophilized (freeze-dried) powder
Purified
Reconstitution:
Restore in sterile 0.4% NaHCO3
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