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.
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
These...
Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...

You might also read

Related Articles

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

Sort by
Same author

First-in-Human Study of a Novel Coronary Crossing Device for Calcified Lesions.

JACC. Cardiovascular interventions·2025
Same author

Analysis of seat belt buckle release forces in post-rollover scenarios: Implications for passenger safety.

Applied ergonomics·2025
Same author

A fatigue failure framework for the assessment of highly variable low back loading using inertial motion capture - a case study.

Ergonomics·2025
Same author

Critical review of chronic toxicity testing approaches with the saltwater mysid (Americamysis bahia) used in pesticide registration.

Environmental toxicology and chemistry·2025
Same author

Scan/rescan reliability of magnetic resonance imaging (MRI).

European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society·2025
Same author

Expedited conveyance of out-of-hospital-cardiac arrest patients with STEMI and shockable rhythms to Cardiac Arrest Centres - A feasibility pilot study of the British Cardiovascular Intervention Society conveyance algorithm.

Resuscitation·2025
Same journal

Nondenaturing Polyacrylamide Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Purification and Concentration of DNA from Aqueous Solutions: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Expression of Proteins Using Semliki Forest Virus Vectors: Protein Expression.

Current protocols in molecular biology·2021
Same journal

Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions: DNA-Protein Interactions.

Current protocols in molecular biology·2021
Same journal

Separation of Double- and Single-Stranded Nucleic Acids Using Hydroxylapatite Chromatography: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Pulsed-Field Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
See all related articles

Related Experiment Video

Updated: Jul 3, 2026

Immunoblot Analysis
16:01

Immunoblot Analysis

Published on: June 20, 2008

Immunoblotting and immunodetection.

Sean Gallagher1, Scott E Winston, Steven A Fuller

  • 1UVP, Inc., Upland, California, USA.

Current Protocols in Molecular Biology
|July 18, 2008
PubMed
Summary
This summary is machine-generated.

Western blotting is a key technique for identifying specific antigens using antibodies. This protocol details protein solubilization, separation, membrane transfer, and immunodetection for reliable results.

More Related Videos

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
09:11

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Published on: November 14, 2016

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions
07:16

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Published on: January 5, 2024

Related Experiment Videos

Last Updated: Jul 3, 2026

Immunoblot Analysis
16:01

Immunoblot Analysis

Published on: June 20, 2008

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
09:11

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Published on: November 14, 2016

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions
07:16

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Published on: January 5, 2024

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Immunoblotting, commonly known as western blotting, is a widely used technique.
  • It enables the identification of specific protein targets.
  • This process relies on the interaction between antigens and antibodies.

Purpose of the Study:

  • To provide comprehensive protocols for immunoblotting.
  • To cover all essential steps from sample preparation to detection.
  • To ensure reliable and reproducible experimental outcomes.

Main Methods:

  • Protein sample solubilization using SDS and reducing agents.
  • Separation of proteins via SDS-PAGE.
  • Electrophoretic transfer of separated proteins to a membrane.
  • Immunodetection using primary and secondary antibodies.
  • Detection via chromogenic or luminescent substrates.
  • Optional membrane stripping and reprobing.

Main Results:

  • Successful identification of specific antigens.
  • Demonstration of the complete immunoblotting workflow.
  • Validation of protocols for various experimental needs.

Conclusions:

  • Immunoblotting is a versatile and powerful method for protein analysis.
  • Standardized protocols are crucial for accurate antigen identification.
  • The described methods facilitate robust western blotting experiments.