Analysis Biomolecular Interactions of PaParE and PaParD by BLI (CAT#: STEM-MB-0184-CJ)

Introduction

Type II toxin–antitoxin (TA) systems, comprised of two non-secreted tightly interacting proteins, are encoded on plasmids as well as in bacterial and archaeal chromosomes.

The ParE class of toxin, which is present throughout the Firmicutes and alpha- and gamma-Proteobacteria phyla. Among the most prevalent toxin families is the RelE/ParE superfamily, which contains a conserved protein fold but, as typical for TA systems, has low sequence conservation. The ParE toxins comprise a subfamily that has been demonstrated to inhibit gyrase-mediated supercoiling in vitro. In addition, the chromosomal location of numerous ParDE operons raises questions about their roles in bacterial physiology. For the ParDE2 system from Mycobacterium tuberculosis, was noted in cells expressing the ParE toxin, has a protective rather than toxic effect.

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Principle Applications Procedure Materials Notes Related Products

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Immunology/Inflammation, Toxicology

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.

Materials

• Equipment: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules

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