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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.

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...

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Related Experiment Video

Updated: Aug 29, 2025

Author Spotlight: Advancing SERS Technology: Au@Carbon Dot Nanoprobes for Label-Free Analysis and Imaging

Author Spotlight: Advancing SERS Technology: Au@Carbon Dot Nanoprobes for Label-Free Analysis and Imaging

Published on: June 9, 2023

Core-shell SERS nanotags-based western blot.

Bing Liu¹, Hanyu Tang¹, Qian Liu¹

¹Medical School, Institute of Reproductive Medicine, Nantong University, Nantong, 226001, China.

|September 10, 2022

Summary

This summary is machine-generated.

This study introduces surface-enhanced Raman scattering-based Western blot (SERS-WB) for highly sensitive protein detection. SERS-WB offers improved quantification of low-abundance proteins, advancing life science research.

Keywords:

Gold-silver core-shell nanoparticle Protein identification Quantitative detection SERS nanotag Western blot

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Area of Science:

Biotechnology
Analytical Chemistry
Molecular Biology

Background:

Western blot (WB) is a standard technique for protein identification.
Accurate quantification of low-abundance proteins remains a significant challenge in WB.
Existing methods lack the sensitivity required for detecting trace protein levels.

Purpose of the Study:

To develop a novel surface-enhanced Raman scattering-based Western blot (SERS-WB) method.
To enhance the sensitivity and accuracy of protein detection, particularly for low-abundance proteins.
To simplify the Western blot procedure through direct colorimetric signal identification.

Main Methods:

Development of SERS nanotags using gold-silver core-shell nanoparticles with Nile blue A (NBA) molecules.
Application of SERS nanotags as quantitative labels for proteins in Western blot.
Utilizing nanoparticle aggregation for direct colorimetric identification of target proteins on nitrocellulose membranes.

Main Results:

Achieved excellent sensitivity for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein with a detection limit of 0.15 pg.
Demonstrated a wide linear dynamic range (LDR) from 382 fg to 382 ng.
Enabled direct colorimetric identification of target proteins, simplifying the experimental workflow.

Conclusions:

The novel SERS-WB strategy significantly improves protein detection sensitivity and quantification.
This technique offers a simplified and highly sensitive alternative to traditional Western blotting.
The proposed method provides innovative approaches for technological advancements in Western blot applications.