Related Experiment Video
Updated: Aug 14, 2026
22:45
Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
Published on: August 22, 2007
Biotinylated proteins as molecular weight standards on Western blots
Analytical Biochemistry
|February 1, 1986
Summary
Biotinylated protein molecular weight standards are easily prepared and effective for detecting proteins via Western blotting. These standards simplify protein analysis in various immunodetection procedures.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Analysis
Background:
- Accurate molecular weight determination is crucial for protein identification and characterization.
- Traditional protein molecular weight standards may have limitations in certain detection methods.
Purpose of the Study:
- To develop easily prepared and highly effective biotinylated protein molecular weight standards.
- To demonstrate the utility of these standards in protein detection assays.
Main Methods:
- Biotinylation of protein standards using biotinyl-N-hydroxysuccinimide ester.
- Separation of biotinylated proteins via SDS-PAGE.
- Transfer to nitrocellulose membranes.
- Detection using a streptavidin-biotin/horseradish peroxidase complex and 4-chloro-1-naphthol/hydrogen peroxide substrate.
Main Results:
- Successfully prepared biotinylated protein molecular weight standards.
- Demonstrated effective detection of biotinylated protein bands on nitrocellulose membranes.
- Confirmed the compatibility of these standards with common immunodetection protocols.
Conclusions:
- Biotinylated protein standards are readily synthesized.
- These standards offer a reliable and versatile tool for molecular weight estimation in protein analysis.
- They are particularly useful for immunodetection techniques involving nitrocellulose transfers.
More Related Videos
Related Concept Videos
01:27SDS-PAGE
Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
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.