OmpA

Outer Membrane Protein-A Bacterial Recombinant
Cat. No.
BT2392
Source
Escherichia Coli.
Synonyms
Outer Membrane Protein-A, OmpA.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity

Greater than 98.0% as determined by:
(a) Analysis by Gel filtration.
(b) Analysis by SDS-PAGE.

Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
Shipped with Ice Packs
In Stock
Quick Inquiry

Description

The recombinant form was found to be undistinguishable from the wild type when examined by SDS-PAGE and gel filtration chromatography yielding a 50.5 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.
The OmpA is purified by proprietary chromatographic techniques.

Product Specs

Introduction
OmpA is a key outer membrane protein found in many Gram-negative bacteria, including A.salmonicida, Shigella dysenteriae, and E.coli. It plays a crucial role in maintaining cell structure, morphology, and porin activity. Additionally, OmpA contributes to bacterial conjugation and acts as a receptor for bacteriophages. Achromogenic atypical Aeromonas salmonicida, the bacterium responsible for goldfish ulcer disease, relies on a paracrystalline outer membrane A-layer protein for its virulence. To study this protein, the gene encoding the monomeric form of A-protein from this bacterium was cloned into a pET-3d plasmid. This allowed for the expression and production of recombinant A-protein in E.coli BL21(DE3). After expression, the protein was isolated from inclusion bodies using a solubilization-renaturation method and further purified to over 95% purity by ion exchange chromatography on Q-Sepharose. This purified recombinant A-protein was then compared to A-protein directly isolated from atypical A.salmonicida using biochemical, immunological, and molecular techniques.
Description
The recombinant A-protein was found to be very similar to the naturally occurring (wild type) form. Both SDS-PAGE and gel filtration chromatography showed it to be a 50.5 kDa monomer. Immunological techniques, including ELISA and Western Blot using polyclonal and monoclonal antibodies, further confirmed the similarity between the recombinant and wild type A-proteins. Significantly, all forms of A-protein were able to activate the release of tumor necrosis factor alpha from murine macrophages. For further details, refer to Maurice et al. (1999) Protein Expression and Purification 16, 396-404. The purification of OmpA is carried out using proprietary chromatographic methods.
Physical Appearance
Sterile Filtered White lyophilized powder
Formulation
The OmpA protein is provided as a lyophilized powder. The protein was initially in a 1mg/ml solution with 0.02% NaHCO3 before lyophilization.
Solubility
To reconstitute the lyophilized OmpA, it is recommended to use sterile 0.4% NaHCO3. The reconstituted solution should have a minimum concentration of 100µg/ml. This solution can be further diluted with other aqueous solutions if needed.
Stability
Lyophilized Bacterial Outer Membrane Protein-A is stable at room temperature for up to 3 weeks. However, for long-term storage, it is best to store it in a dry environment below -18°C. After reconstitution, the OmpA solution should be stored at 4°C for short-term use (2-7 days) and at -18°C for long-term storage. For long-term storage, adding a carrier protein like 0.1% HSA or BSA is recommended. Repeated freezing and thawing of the protein should be avoided.
Purity
The purity of the OmpA is greater than 98.0%. This is determined using the following methods: (a) Gel filtration analysis. (b) SDS-PAGE analysis.
Biological Activity
Experiments were carried out to investigate how bacterial and recombinant A-layer protein interact with murine macrophages, particularly their impact on the internal processes of primed macrophages. This was achieved by exposing peritoneal macrophages to latex beads coated with A-protein and measuring the resulting cytotoxic product. Macrophages derived from thioglycolate stimulation displayed a baseline level of activation (18% cytotoxicity), which significantly increased (48% cytotoxicity) in the presence of latex beads. When the latex beads were coated with any of the three A-protein variants, the cytotoxicity level increased even further, from 48% to 91% (mean +/- SEM).
Synonyms
Outer Membrane Protein-A, OmpA.
Source
Escherichia Coli.
Amino Acid Sequence
mdvvispndn tfvttslasv tkqpvldfst aqqnltlnfs evgdlknngf ivleiqgegq fndaeirqwl sngfwrrpft gllvnpndhg nfansgevnd vrkffkiisd gtqltivhti dsngkrlrla lasdveetin fadaevelkl nlanqafklt sgsqgtvalt agalwnasyt adpvatkplf klgklfqlsl tnagkatalv segflklnig danisatdfa itnvttnqti qrdkvnltlt gdvsafkkda ngnlvnkaga sigwkaaadg qsatavlgag nmaggvqnal aafgtlyvaa dntvpvpavn fnvkaeiqgd sqatynyfkd eladlfiltr dgmkfdtitt gttsanlihi rdvsnilpte ggkifvtite yadhaangrg egtvlvtrka lsvtlpsgga vtlkpadvaa dvgasitagr qarlvfevet nqgevavkks naegvdiqng trgtaplvdf tl.

Product Science Overview

Introduction

Outer Membrane Protein-A (OmpA) is a significant protein found in the outer membrane of Gram-negative bacteria, such as Escherichia coli (E. coli). It plays a crucial role in maintaining the structural integrity of the bacterial cell and is involved in various functions, including bacterial adhesion, biofilm formation, and immune evasion .

Historical Context

OmpA was first identified as a heat-modifiable protein in E. coli in 1974 and was originally purified in 1977 . Since then, it has been extensively studied due to its importance in bacterial physiology and its potential as a target for therapeutic interventions.

Structure and Function

OmpA is a porin protein with a molecular mass ranging from 28 kDa to 36 kDa . It is a surface-exposed protein that forms a beta-barrel structure, allowing it to function as a channel for the passage of small molecules. OmpA is also involved in interactions with host cells, contributing to the pathogenicity of certain bacteria, such as Acinetobacter baumannii .

Recombinant OmpA

Recombinant OmpA refers to the protein produced through recombinant DNA technology. This involves cloning the gene encoding OmpA into an expression vector, which is then introduced into a host organism, typically E. coli. The host organism expresses the OmpA protein, which can be purified for various applications.

Applications
  1. Vaccine Development: Recombinant OmpA has been explored as a potential vaccine candidate due to its ability to induce a protective immune response against bacterial infections .
  2. Therapeutic Target: OmpA is considered a potential therapeutic target for treating infections caused by multidrug-resistant bacteria, such as A. baumannii .
  3. Biotechnological Applications: OmpA can be used in bacterial surface display systems to expose heterologous proteins or peptides on the bacterial surface for applications like peptide library screening and live bacterial vaccine design .

Quick Inquiry

Personal Email Detected
Please use an institutional or corporate email address for inquiries. Personal email accounts ( such as Gmail, Yahoo, and Outlook) are not accepted. *

Category

Service

Related Products

Omp Pylori
Helicobacter Pylori Outer Membrane Protein Recombinant
Cat. No.
BT19504
Source
E.Coli
THE BIOTEK
THE BioTek is a global biotech company offering highly purified proteins for research in cancer, apoptosis, development, endocrinology, immunology, neuroscience, proteases, and stem cells.
  • Contact
  • Address: 1925 E Maple Ave, El Segundo, CA 90245 United States
  • Phone : +1 201-285-7826
  • Email : info@thebiotek.com
  • Hours : Mon.-Fri. 9:00 AM - 5:00 PM
© Copyright 2024 Thebiotek. All Rights Reserved.