Protein identification on archived 2-D gels
Hiroyuki Matsumoto1, Nobuaki Takemori, Naoka Komori
1University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. hiroyuki-matsumoto@ouhsc.edu
Methods in Molecular Biology (Clifton, N.J.)
|May 16, 2012
Summary
Researchers can now identify old, archived proteins using modern genome and proteomics data. This protocol helps uncover the identity of proteins isolated decades ago when identification methods were limited.
Area of Science:
- Biochemistry
- Proteomics
- Genomics
Background:
- Proteins isolated decades ago via two-dimensional gel electrophoresis were archived without definitive identification.
- Limited analytical techniques at the time of isolation prevented protein characterization.
Purpose of the Study:
- To provide a protocol for identifying previously uncharacterized proteins from historical archives.
- To leverage current genomic and proteomic data for retrospective protein identification.
Main Methods:
- Utilizing available genome information.
- Applying modern proteomics techniques.
- Cross-referencing archived protein data with current databases.
Main Results:
- Successful identification of long-archived, uncharacterized proteins is now feasible.
- The described protocol enables retrospective analysis of historical protein samples.
Conclusions:
- Modern genomic and proteomic advancements allow for the identification of historical protein samples.
- This protocol empowers researchers to revisit and identify proteins from past experiments.
Related Concept Videos
01:22Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
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.