Peptide Mass Fingerprinting Using MALDI-TOF Mass Spectrometry
Large-format 2D gel electrophoresis systems in routine operation are capable of resolving several thousand cellular proteins in 1 or 2 d (1,2). For the last decade, a combination of Edman microsequence analysis and identification of proteins by staining with specific antibodies has been used to systematically identify proteins and establish cellular databases (3, 4, 5). There are, however, significant problems associated with these approaches. Most proteins are only present in the low-to upper-femtomole range, which is significantly below the level at which automated sequencers can reliably operate (6,7). The relatively slow speed of the Edman process also means that the number of proteins is too great to permit large-scale characterization within any useful period of time. The use of monoclonal antibodies, while both rapid and sensitive, requires the ready availability of a large pool of specific antibody probes.
KeywordsSyringe Lysine Bicarbonate Trypsin MeOH
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