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

Western Blotting01:15

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
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...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
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...

You might also read

Related Articles

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

Sort by
Same author

Nuclear Proteomics to Understand the Promotive Effect of Plant-Derived Smoke Solution on Wheat Under Salt Stress.

Proteomes·2026
Same author

Analyzing the Specificity of KAWLR Genetic Resources in Afghan Landrace Wheat for Ca-Rich High pH Soil Tolerance Using Proteomics.

International journal of molecular sciences·2026
Same author

Green Synthesized Titanium Oxide Nanoparticles Promote Salt Tolerance in Soybean.

International journal of molecular sciences·2025
Same author

Role of Plant-Derived Smoke Solution on Plants Under Stress.

International journal of molecular sciences·2025
Same author

The Growth of Soybean (<i>Glycine max</i>) Under Salt Stress Is Modulated in Simulated Microgravity Conditions.

Cells·2025
Same author

Protein-Based Mechanism of Wheat Growth Under Salt Stress in Seeds Irradiated with Millimeter Waves.

International journal of molecular sciences·2025

Related Experiment Video

Updated: Jun 23, 2026

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
09:45

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

Published on: January 29, 2018

Western blotting/Edman sequencing using PVDF membrane.

Setsuko Komatsu1

  • 1National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba, 305-8518, Japan. skomatsu@affrc.go.jp

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed
Summary
This summary is machine-generated.

Western blotting and Edman sequencing on polyvinylidene difluoride (PVDF) membranes are popular for protein primary structure determination. This method, often combined with 2D-PAGE, effectively sequences complex protein mixtures.

More Related Videos

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology
05:59

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
22:45

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Published on: August 22, 2007

Related Experiment Videos

Last Updated: Jun 23, 2026

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
09:45

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

Published on: January 29, 2018

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology
05:59

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
22:45

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Published on: August 22, 2007

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein primary structure determination is crucial for understanding protein function.
  • Western blotting and Edman sequencing are established techniques for this purpose.
  • Polyvinylidene difluoride (PVDF) membranes are commonly used for protein transfer and sequencing.

Purpose of the Study:

  • To describe the western blotting/Edman sequencing technique using PVDF membranes.
  • To highlight its application in determining the primary structure of proteins.
  • To present a combined approach with 2D-PAGE for complex protein mixtures.

Main Methods:

  • Protein samples are separated by SDS-polyacrylamide gel electrophoresis (PAGE).
  • Proteins are transferred to a PVDF membrane via electroblotting.
  • The PVDF membrane with the protein is subjected to Edman sequencing.
  • Deblocking solutions and alternative methods like the Cleveland method are used if sequencing fails.

Main Results:

  • The western blotting/Edman sequencing technique is a popular method for protein primary structure determination.
  • Direct sequencing on PVDF membranes is feasible.
  • Combined use of 2D-PAGE, western blotting, and Edman sequencing enhances sequence determination for crude protein mixtures.
  • This integrated approach overcomes limitations of conventional purification methods.

Conclusions:

  • Western blotting/Edman sequencing on PVDF membranes is a robust technique for protein sequencing.
  • The combination with 2D-PAGE significantly improves the analysis of complex protein samples.
  • This method offers an effective strategy for sequence determination when conventional purification is challenging.