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Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as  cells...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
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...
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...

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Separating Beads and Cells in Multi-channel Microfluidic Devices Using Dielectrophoresis and Laminar Flow
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High electric field strength two-dimensional peptide separations using a microfluidic device.

W Hampton Henley1, J Michael Ramsey

  • 1Department of Chemistry, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Electrophoresis
|September 12, 2012
PubMed
Summary
This summary is machine-generated.

New instrumentation enhances microfluidic 2D separations using capillary electrophoresis (CE) with higher voltages. This boosts resolution, efficiency, and speed, achieving over 4000 peak capacity and rapid peptide detection.

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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
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Published on: October 24, 2018

Area of Science:

  • Analytical Chemistry
  • Separation Science
  • Microfluidics

Background:

  • Previous microfluidic 2D separations were limited by low electric field strength (approx. 200 V/cm).
  • Existing instrumentation restricted maximum potential difference to 8.4 kV, impacting separation performance.

Purpose of the Study:

  • To develop new instrumentation for improved microfluidic 2D separations.
  • To enhance resolution, efficiency, and speed by coupling micellar electrokinetic chromatography (MEKC) with high field strength CE.
  • To overcome limitations of previous instrumentation by employing higher voltages.

Main Methods:

  • Developed a novel circuit combining a high voltage supply with a rapidly switching lower voltage supply.
  • Implemented microfluidic chips with longer channels.
  • Utilized high field strength capillary electrophoresis (CE) coupled with MEKC.
  • Applied voltages exceeding 20 kV for separations.

Main Results:

  • Achieved high electric field strengths in both dimensions, significantly increasing separation resolution, efficiency, and peak capacity.
  • Observed detection rates of up to six peptides per second for a model protein digest.
  • Attained peak capacities exceeding 4000 in some separations using longer channels.
  • Demonstrated a threefold improvement in resolving power compared to low field strength separations.

Conclusions:

  • The new instrumentation enables faster and more powerful microfluidic 2D separations.
  • High field strength CE coupled with MEKC offers superior performance for complex sample analysis.
  • The developed system significantly advances the capabilities of microfluidic separation techniques.