Application data
- Protein identification in complex mixtures is pivotal in the field of proteomics. A common approach comprises the separation of a cell extract by 2D-gel electrophoresis, excision and in-gel enzymatic digestion of a gel spot followed by identification using mass spectrometry.
- Limitations to this approach include restrictions on the range of amenable proteins, together with difficulties arising from the differential detectability of proteolytic peptides. Coupling mass spectrometry with the chromatographic separation of proteolytic peptides from an in-gel digest of a 1D SDS PAGE protein band appears attractive for alleviating these problems.
- Here, we evaluate the combination of single stage protein separation with LC-MALDI-TOF-MS of tryptic digests, in which eluate fractions are spotted automatically onto the MALDI target, and compare results from the conventional gel approach and those obtained using the LC-MALDI-TOF technique.
|
|
| |
 |
|
Comparative study of protein identification by off-line LC-MALDI-MS and 1D-gel electrophoresis MALDI-MS |
 |
| |
Presented at ASMS 2004.
Helen Montgomery1, Sue Francis2, Sadanori Sekiya3, Simon J. Gaskell2, Koichi Tanaka3.
1 Koichi Tanaka MS Research Laboratory, Shimadzu, Manchester, UK;
2 Michael Barber Centre for Mass Spectrometry, UMIST, Manchester, UK;
3 Koichi Tanaka MS Research Laboratory, Shimadzu, Kyoto, Japan.
|
|
