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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.
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...
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...
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...
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,...

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Related Experiment Video

Updated: Jun 4, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

SDS -PAGE and Western Blotting Techniques.

C Blancher1, A Jones

  • 1Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, UK.

Methods in Molecular Medicine
|February 23, 2011
PubMed
Summary

Western blotting, or immunoblotting, identifies specific protein antigens in complex mixtures using antibodies. This technique determines antigen presence, quantity, molecular weight, and extraction efficiency.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Western blotting, or immunoblotting, is a key technique for protein analysis.
  • It allows for the identification of specific antigens within complex protein mixtures.

Purpose of the Study:

  • To outline the principles and stages of immunoblotting.
  • To highlight the importance of sample preparation for accurate protein analysis.

Main Methods:

  • Protein extraction and quantification.
  • Separation of proteins using SDS-PAGE.
  • Transfer to a membrane, blocking, antibody incubation, and detection.

Main Results:

  • Immunoblotting can determine antigen presence, quantity, and molecular weight.

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Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

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Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
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Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

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Last Updated: Jun 4, 2026

Western Blotting: Sample Preparation to Detection
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Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture
09:37

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

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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

  • Sample preparation methods influence extraction efficiency and protein separation.

    • Conclusions:

    • Immunoblotting is a versatile method for characterizing protein antigens.
    • Optimized sample preparation is crucial for successful immunoblotting outcomes.