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Related Concept Videos

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: Apr 11, 2026

Western Blotting: Sample Preparation to Detection

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Post-staining electroblotting for efficient and reliable peptide blotting.

Der-Yen Lee¹, Geen-Dong Chang

¹Graduate Institute of Biochemical Sciences, National Taiwan University, No. 1, Section 4, Roosevelt Road, Taipei, 106, Taiwan.

Methods in Molecular Biology (Clifton, N.J.)

|June 6, 2015

Summary

Post-staining electroblotting effectively transfers stained peptides, including small ones, onto membranes. This method enhances sensitivity and enables further analysis like sequencing and blotting assays.

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Last Updated: Apr 11, 2026

Western Blotting: Sample Preparation to Detection

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Identification of Post-translational Modifications of Plant Protein Complexes

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Published on: February 22, 2014

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Published on: April 10, 2014

Area of Science:

Biochemistry
Proteomics
Analytical Chemistry

Background:

Post-staining electroblotting is established for transferring Coomassie blue-stained proteins.
Conventional electroblotting has limitations in sensitivity and applicability for smaller molecules.

Purpose of the Study:

To evaluate the efficacy of post-staining electroblotting for transferring stained peptides.
To demonstrate the method's suitability for small peptides and subsequent analyses.

Main Methods:

Utilizing post-staining electroblotting to transfer stained peptides from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes.
Comparing the sensitivity of post-staining electroblotting with conventional methods for peptide visualization.

Main Results:

Efficient and reliable transfer of stained peptides, including those below 2 kDa (e.g., bacitracin, granuliberin R).
Post-staining electroblotting is approximately 16-fold more sensitive for insulin visualization compared to conventional electroblotting.
Enhanced peptide retention on PVDF membranes due to selective staining.

Conclusions:

Post-staining electroblotting is a viable and accessible method for peptide blotting.
The technique facilitates further applications such as blot overlay detection, immunoblotting, and N-terminal sequencing.
This method allows for normalized peptide blotting for identification in complex samples.