Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Southern Blot

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...

Western Blotting

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.

You might also read

Related Articles

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

Sort by

Same author

Kaposi's Sarcoma-Associated Herpesvirus Utilizes and Manipulates RNA N<sup>6</sup>-Adenosine Methylation To Promote Lytic Replication.

Journal of virology·2017

Same author

NOTE FROM THE EDITOR.

RNA (New York, N.Y.)·2016

Same author

Erratum: Preparation of Nuclear Extracts from HeLa Cells.

Cold Spring Harbor protocols·2016

Same author

Cotranslational microRNA mediated messenger RNA destabilization.

eLife·2016

Same author

Preparing Cellular DNA from Nuclei or Whole Cells.

Cold Spring Harbor protocols·2015

Same author

Removal of rRNA from Deproteinized, Phenol-Extracted Total RNA by Hybrid Selection.

Cold Spring Harbor protocols·2015

Same journal

Emerging Trends in Mass Spectrometry-Based Quantitative Proteome and Phosphoproteome Profiling in Maize.

Cold Spring Harbor protocols·2026

Same journal

Sample Preparation for Quantitative Proteome and Phosphoproteome Profiling of Maize Tissues.

Cold Spring Harbor protocols·2026

Same journal

High-Throughput Microbial Assay for Amino Acid Measurement in Ground Maize Seed Samples Utilizing Auxotrophic <i>E. coli</i>.

Cold Spring Harbor protocols·2025

Same journal

Grain Quality in Maize.

Cold Spring Harbor protocols·2025

Same journal

High-Throughput Assay for Measuring Phytate and Available Phosphorus in Ground Maize Seed Samples.

Cold Spring Harbor protocols·2025

Same journal

Functional Genomic Analysis of Transposon Insertion Mutant Maize Plants from the UniformMu National Public Resource.

Cold Spring Harbor protocols·2025

See all related articles

Search research articles

Related Experiment Video

Updated: Apr 16, 2026

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

An alternative method for processing northern blots after capillary transfer.

Timothy W Nilsen

Cold Spring Harbor Protocols

|March 4, 2015

Summary

This summary is machine-generated.

This protocol optimizes northern blotting by using a simplified Church and Gilbert hybridization buffer. It effectively minimizes background noise and maximizes specific RNA-DNA hybridization for clear results.

More Related Videos

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Published on: August 22, 2007

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Published on: August 21, 2014

Related Experiment Videos

Last Updated: Apr 16, 2026

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

Published on: August 22, 2007

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Published on: August 21, 2014

Area of Science:

Molecular Biology
Biochemistry

Background:

Northern blotting is a key technique for RNA analysis.
Variations in prehybridization, hybridization, and washing steps exist across laboratories.
Optimizing these steps is crucial for accurate results.

Purpose of the Study:

To present a simplified northern blotting protocol.
To minimize the number of ingredients in solutions.
To reduce background noise and enhance specific hybridization.

Main Methods:

Prehybridization with Church and Gilbert hybridization buffer.
Hybridization of immobilized RNA to a specific DNA probe.
Washing of the membrane followed by autoradiography or phosphorimaging.

Main Results:

The protocol effectively blocks nonspecific probe-binding sites.
Minimized background adherence of probe to membrane.
Maximized specific hybridization to immobilized RNAs.

Conclusions:

This protocol offers an efficient alternative for northern blotting.
It achieves high specificity and low background with fewer reagents.
Suitable for researchers seeking simplified yet effective RNA analysis.