Related Concept Videos
01:15Western 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.
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.
02:57Southern 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...
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...
You might also read
Related Articles
Articles linked to this work by shared authors, journal, and citation graph.
Sort by
Same author
Anti-nuclear antibody screening using HEp-2 cells.
Journal of visualized experiments : JoVEยท2014
Related Experiment Video
Updated: Jun 5, 2026
07:45
Western Blotting: Sample Preparation to Detection
Published on: October 14, 2010
Western blotting: sample preparation to detection.
1Research and Development, EMD Chemicals Inc.
Journal of Visualized Experiments : Jove
|December 30, 2010
Summary
Western blotting is a key protein detection method. This technique uses gel electrophoresis and antibody probing to identify specific proteins in biological samples.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- Western blotting is a widely used analytical technique.
- It is essential for detecting specific proteins in complex biological samples.
- Understanding the methodology is crucial for accurate protein analysis.
Purpose of the Study:
- To describe the fundamental principles of Western blotting.
- To outline the key steps involved in protein separation, transfer, and detection.
- To highlight the versatility of the technique in various research contexts.
Main Methods:
- Utilizes gel electrophoresis (denaturing or native conditions) to separate proteins based on polypeptide length or 3-D structure.
- Involves transferring separated proteins onto a membrane, such as nitrocellulose or PVDF.
Main Results:
- Successful identification and quantification of specific proteins within a sample.
- Provides insights into protein expression levels and molecular weight.
- Confirms the presence or absence of target proteins.
Conclusions:
- Western blotting is a robust and sensitive method for specific protein detection.
- The technique's multi-step process ensures high specificity and reliability.
- It remains an indispensable tool in biological and medical research for protein analysis.