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

22.2K
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.
22.2K
Southern Blot02:57

Southern Blot

24.6K
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...
24.6K
Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

8.3K
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...
8.3K
SDS-PAGE01:27

SDS-PAGE

36.2K
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...
36.2K
Electrophoresis: Overview01:20

Electrophoresis: Overview

5.1K
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...
5.1K

You might also read

Related Articles

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

Sort by
Same author

VP2-targeted sandwich ELISA (sELISA) enables direct detection of Senecavirus A (SVA).

Journal of virology·2026
Same author

When Inhibition Fails: Adolescent Nicotine Disrupts Opioid Control of Ventral Tegmental Area GABA Neurons.

Biological psychiatry·2026
Same author

GABA-glutamate corelease is a mechanism for state-dependent neurotransmission.

bioRxiv : the preprint server for biology·2025
Same author

Mu-opioid receptor activation potentiates excitatory transmission at the habenulo-peduncular synapse.

Cell reports·2025
Same author

Parkinson's Disease-vulnerable and -resilient dopamine neurons display opposite responses to excitatory input.

bioRxiv : the preprint server for biology·2025
Same author

Ventral tegmental area interneurons revisited: GABA and glutamate projection neurons make local synapses.

eLife·2025
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Apr 7, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

147.6K

The Western Blot.

Thomas S Hnasko1, Robert M Hnasko

  • 1Department of Neurosciences, School of Medicine, University of California San Diego (UCSD), 9500 Gilman Drive (BRF2), La Jolla, CA, 92093, USA, thnasko@ucsd.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 11, 2015
PubMed
Summary
This summary is machine-generated.

Western blotting is a key protein analysis technique. It uses gel electrophoresis, membrane transfer, and antibody detection for identifying specific proteins and their molecular weights.

More Related Videos

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

39.1K
The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis
11:43

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

28.1K

Related Experiment Videos

Last Updated: Apr 7, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

147.6K
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

39.1K
The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis
11:43

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

28.1K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Western blotting is a standard method for protein analysis.
  • It relies on specific antibody-antigen interactions.
  • This technique allows for protein identification and molecular weight determination.

Purpose of the Study:

  • To outline the essential steps of Western blotting.
  • To provide a guide for qualitative and semiquantitative protein identification.
  • To detail the process from sample preparation to immunodetection.

Main Methods:

  • Protein separation using gel electrophoresis.
  • Transfer of separated proteins to a membrane support.
  • Selective immunodetection of immobilized target antigens.

Main Results:

  • Successful qualitative and semiquantitative identification of specific proteins.
  • Accurate determination of protein molecular weights within complex mixtures.
  • Demonstration of the utility of antibody-antigen specificity in protein analysis.

Conclusions:

  • Western blotting is a versatile and crucial technique in molecular biology.
  • The outlined steps provide a foundational understanding for performing Western blots.
  • This method is indispensable for validating protein expression and characterization.