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Related Concept Videos

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...
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,...
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...
SDS-PAGE01:27

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed  polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...

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Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
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Published on: February 24, 2011

Supported bilayer electrophoresis under controlled buffer conditions.

Christopher F Monson1, Hudson P Pace, Chunming Liu

  • 1Department of Chemistry, Texas A&M University, 3255 TAMU, College Station, Texas 77843, United States.

Analytical Chemistry
|February 16, 2011
PubMed
Summary

A novel flow cell device enables controlled electrophoretic movement of lipids and proteins in supported phospholipid bilayers. This system allows independent manipulation of electrophoretic and electroosmotic forces, crucial for biomembrane studies.

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Area of Science:

  • Biophysics
  • Materials Science
  • Biochemistry

Background:

  • Supported phospholipid bilayers are crucial models for cell membranes.
  • Studying the movement of lipids and proteins within bilayers is essential for understanding membrane dynamics.
  • Existing methods often struggle with pH stability and separation of electrophoretic forces.

Purpose of the Study:

  • To develop a pH-controlled flow cell for precise electrophoretic manipulation of charged molecules in supported phospholipid bilayers.
  • To investigate the electrophoretic mobility of lipids and membrane-associated proteins under varying pH and buffer conditions.
  • To differentiate and control electrophoretic and electroosmotic contributions to macromolecular transport.

Main Methods:

  • Construction of a pH-controlled flow cell device that isolates electrolysis products.
  • Utilizing supported phospholipid bilayers with charged lipids and membrane-associated proteins.
  • Employing dye-conjugated lipids (Texas Red-DHPE) and streptavidin bound to biotinylated lipids for mobility studies.
  • Manipulating buffer pH, ionic strength, and incorporating polyethylene glycol (PEG) cushions.

Main Results:

  • The flow cell maintained stable pH (±0.2 units) for extended periods at up to 10 mM salt concentrations.
  • Texas Red-DHPE showed constant electrophoretic mobility across a wide pH range (3.3-9.3).
  • Streptavidin exhibited pH-dependent migration, shifting from cathodic to anodic movement, indicating charge modulation.
  • PEG cushions and increased ionic strength reduced electroosmotic forces on streptavidin without significantly affecting lipid mobility.

Conclusions:

  • The developed flow cell provides a stable environment for studying electrophoresis in supported phospholipid bilayers.
  • Protein charge modulation significantly influences electrophoretic mobility in response to pH changes.
  • Electrophoretic and electroosmotic forces can be independently controlled by adjusting buffer conditions and using PEG cushions, enabling precise manipulation of biomolecules within bilayers.