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Related Concept Videos

Western Blotting01:15

Western Blotting

22.3K
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.
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Southern Blot02:57

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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...
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Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

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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...
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Electrophoresis: Overview01:20

Electrophoresis: Overview

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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...
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SDS-PAGE01:27

SDS-PAGE

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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...
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Related Experiment Video

Updated: Apr 11, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

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Western blotting: an introduction.

Biji T Kurien1, R Hal Scofield

  • 1Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA, biji-kurien@omrf.org.

Methods in Molecular Biology (Clifton, N.J.)
|June 6, 2015
PubMed
Summary
This summary is machine-generated.

Western blotting is a key technique for detecting proteins, especially those present in low amounts. This method transfers protein patterns from a gel to a membrane, with various transfer techniques available since 1979.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Western blotting is a crucial technique for protein immunodetection.
  • It is particularly valuable for identifying low-abundance proteins.

Purpose of the Study:

  • To highlight the significance of protein transfer in Western blotting.
  • To provide an overview of the evolution of protein transfer methods.

Main Methods:

  • Protein transfer from gel to microporous membrane.
  • Electrophoretic and non-electrophoretic transfer techniques.

Main Results:

  • Protein transfer is fundamental to Western blotting.
  • Multiple methods for protein transfer have been developed since 1979.

Conclusions:

  • Western blotting remains an important procedure for protein analysis.
  • The evolution of transfer methods has increased the versatility of protein blotting.