Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

8.4K
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
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

1.9K
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

36.6K
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...
36.6K
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

2.1K
In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
2.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Performance of preclinical models in predicting drug-induced liver injury in humans: a systematic review.

Scientific reports·2021
Same author

Fluorescent Western Blotting: High Sensitivity Detection of Multiple Targets.

Current protocols in pharmacology·2020
Same author

2-D Western blotting for evaluation of antibodies developed for detection of host cell protein.

Methods in molecular biology (Clifton, N.J.)·2015
Same author

Flexible and accessible workflows for improved proteogenomic analysis using the Galaxy framework.

Journal of proteome research·2014
Same author

The chromosome-centric human proteome project: a call to action.

Journal of proteome research·2012
Same author

Workflow for analysis of high mass accuracy salivary data set using MaxQuant and ProteinPilot search algorithm.

Proteomics·2012

Related Experiment Video

Updated: Apr 15, 2026

Sample Preparation for Mass-spectrometry-based Proteomics Analysis of Ocular Microvessels
11:32

Sample Preparation for Mass-spectrometry-based Proteomics Analysis of Ocular Microvessels

Published on: February 22, 2019

12.0K

In-gel peptide IEF sample preparation for LC/MS analysis.

Tom Berkelman1, Sricharan Bandhakavi, Aran Paulus

  • 1Bio-Rad Laboratories, Life Science Group, Hercules, CA, USA, Thomas_berkelman@bio-rad.com.

Methods in Molecular Biology (Clifton, N.J.)
|March 31, 2015
PubMed
Summary
This summary is machine-generated.

This study presents an improved method for protein analysis using in-gel isoelectric focusing (IEF) with immobilized pH gradient (IPG) strips. This technique enhances proteome coverage in liquid chromatography-mass spectrometry (LC-MS) workflows.

More Related Videos

Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing Frog Embryo
09:18

Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing Frog Embryo

Published on: April 21, 2022

2.3K
Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
08:07

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

9.1K

Related Experiment Videos

Last Updated: Apr 15, 2026

Sample Preparation for Mass-spectrometry-based Proteomics Analysis of Ocular Microvessels
11:32

Sample Preparation for Mass-spectrometry-based Proteomics Analysis of Ocular Microvessels

Published on: February 22, 2019

12.0K
Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing Frog Embryo
09:18

Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing Frog Embryo

Published on: April 21, 2022

2.3K
Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
08:07

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

9.1K

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Proteolytic digestion followed by liquid chromatography-mass spectrometry (LC-MS) is a standard method for protein identification.
  • Isoelectric focusing (IEF) can be employed prior to LC-MS to improve proteome coverage.
  • Existing IEF methods may have limitations in sample preparation or compatibility.

Purpose of the Study:

  • To describe a novel method for in-gel isoelectric focusing (IEF) separation of proteolytic digests.
  • To demonstrate the utility of commercially available immobilized pH gradient (IPG) strips for this application.
  • To enhance the depth of proteome coverage in LC-MS analyses.

Main Methods:

  • Proteolytic digestion of protein samples.
  • In-gel isoelectric focusing (IEF) using immobilized pH gradient (IPG) strips.
  • Subsequent analysis by liquid chromatography-mass spectrometry (LC-MS).

Main Results:

  • Successful implementation of in-gel IEF separation for proteolytic digests.
  • Demonstrated compatibility with standard IPG strips and IEF instrumentation.
  • Potential for increased proteome coverage compared to standard LC-MS.

Conclusions:

  • The described in-gel IEF method offers a practical approach to enhance proteomic analysis.
  • This technique is accessible using widely available commercial materials.
  • The method provides a valuable tool for deeper proteome exploration.