Related Experiment Video

Updated: Apr 21, 2026

Polysome Profiling in Leishmania, Human Cells and Mouse Testis
14:32

Polysome Profiling in Leishmania, Human Cells and Mouse Testis

Published on: April 8, 2018

19.1K

Separation of basic proteins from Leishmania using a combination of Free flow electrophoresis (FFE) and 2D

Marie-Christine Brotherton1, Gina Racine, Marc Ouellette

  • 1Centre de Recherche en Infectiologie, CHU-Québec research centre, 2705 boul Laurier, Québec, Canada, G1V 4G2.

Methods in Molecular Biology (Clifton, N.J.)
|November 13, 2014
PubMed

Insights

This study introduces a new method to better visualize basic proteins from Leishmania using combined Free Flow Electrophoresis in Isoelectric Focusing mode and 2D gel electrophoresis. This technique significantly improves the detection of these underrepresented proteins.

Area of Science:

  • Proteomics
  • Molecular Biology
  • Parasitology

Background:

  • Basic proteins are crucial in intracellular organisms like Leishmania.
  • These proteins are often underrepresented and difficult to analyze using standard 2D gel electrophoresis.
  • Improved visualization techniques are needed for comprehensive Leishmania proteome analysis.

Purpose of the Study:

  • To develop an improved method for the fractionation and visualization of basic proteins from Leishmania.
  • To overcome the limitations of traditional 2D gel electrophoresis for basic protein analysis.
  • To enhance the detection of Leishmania proteins with a basic isoelectric point (pI).

Main Methods:

  • Fractionation of basic proteins using Free Flow Electrophoresis in Isoelectric Focusing mode (IEF-FFE).
  • Subsequent protein separation using two-dimensional gel electrophoresis (2-DE) under basic conditions.
  • Combined application of IEF-FFE and basic 2-DE for Leishmania protein analysis.

Main Results:

  • The combined IEF-FFE and basic 2-DE method effectively fractionates basic proteins.
  • This technique significantly improves the visualization of Leishmania proteins with basic pI.
  • The method addresses the underrepresentation issue of basic proteins on 2D gels.

Conclusions:

  • The combination of IEF-FFE and basic 2-DE is a powerful approach for analyzing basic proteins.
  • This method represents a significant advancement for Leishmania proteome research.
  • Enhanced visualization of basic proteins aids in understanding their roles in Leishmania.

More Related Videos

Electrophoretic Separation of Proteins08:17

Electrophoretic Separation of Proteins

Published on: June 12, 2008

35.7K
Deciphering the Molecular Mechanism and Function of Pore-Forming Toxins Using Leishmania major08:17

Deciphering the Molecular Mechanism and Function of Pore-Forming Toxins Using Leishmania major

Published on: October 28, 2022

2.0K

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...
8.4K
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...
5.3K
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,...
1.9K
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...
37.0K
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
122.6K