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Related Concept Videos

Electrophoresis: Overview01:20

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
Capillary Electrophoresis: Applications01:30

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Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
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Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...

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Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis
08:09

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Published on: January 7, 2017

Measuring aptamer equilibria using gradient micro free flow electrophoresis.

Ryan T Turgeon1, Bryan R Fonslow, Meng Jing

  • 1Department of Chemistry, 207 Pleasant Street South East, University of Minnesota, Minneapolis, Minnesota 55455, USA.

Analytical Chemistry
|April 9, 2010
PubMed
Summary
This summary is machine-generated.

Gradient micro free flow electrophoresis rapidly measured DNA aptamer-target equilibria, improving speed and precision over traditional methods. This technique revealed complex stoichiometries and dissociation constants for IgE and HIVRT targets.

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Primer-Free Aptamer Selection Using A Random DNA Library
11:14

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Published on: July 26, 2010

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Studying DNA aptamer-target interactions is crucial for understanding molecular recognition.
  • Traditional methods like affinity capillary electrophoresis (ACE) can be time-consuming and lack precision.
  • Accurate determination of binding equilibria and stoichiometries is essential for aptamer characterization.

Purpose of the Study:

  • To develop and validate a rapid, high-precision method for observing DNA aptamer-target binding equilibria.
  • To characterize the binding kinetics and stoichiometry of DNA aptamers with IgE and HIVRT targets.
  • To compare the efficiency of micro free flow electrophoresis (muFFE) with existing techniques like ACE.

Main Methods:

  • Utilized gradient micro free flow electrophoresis (muFFE) for real-time equilibrium analysis.
  • Mixed continuous streams of DNA aptamer with increasing concentrations of target ligands (IgE or HIVRT).
  • Separated unbound aptamer from ligand-aptamer complexes within the muFFE device.

Main Results:

  • muFFE enabled measurement of equilibrium distribution across 300 discrete target concentrations in under 5 minutes.
  • Estimated the dissociation constant for the aptamer-IgE complex at 48 +/- 3 nM.
  • Observed and characterized complex stoichiometries for aptamer-HIVRT interactions, including 3:1, 2:1, and 1:1 ratios.

Conclusions:

  • Gradient muFFE offers a significant advancement in speed and precision for aptamer-target interaction analysis.
  • The method allows for detailed characterization of binding affinities and complex formation.
  • muFFE is a powerful tool for high-throughput screening and characterization of aptamer-based systems.