Originally Posted by nduong
Can I apply the above method to 1-D gel electrophoresis?
Remember that bands in 1D gels can and usually consist of several if not dozens of proteins that happen to be the same MW. So you'll most likely be working with a mixture of proteins, not one single novel protein.
As long as the proteins in that band/mixture are present in the same amount then you can perform trypsinization and LC-MS-MS and get a decent list of proteins from that band. And like I said you'll have peptide sequences which can be synthetically made and used for creating unique antibodies. And from there you'll have to find the "right" antibody for something like Immunopurification....Also with enough hits (high confidence peptides) you may get the gene ID and go to recombinant genetics for amplifying the amount of protein....
The drawback is that it can backfire for rare proteins; they may not show up, because their signal or presence can and will be drowned out by the most abundant proteins...so the strength of the technology is to work with the most purified level of protein you can get.