Chemistry, Mass Spectrometry and Peptide-Mass Databases: Evolution of Methods for the Rapid Identification and Mapping of Cellular Proteins

Pappin, D. J. C., Rahman, D., Hansen, H. F., Bartlet-Jones, M., Jeffery, W., Bleasby, A. J. (1996) Chemistry, Mass Spectrometry and Peptide-Mass Databases: Evolution of Methods for the Rapid Identification and Mapping of Cellular Proteins. In: Mass Spectrometry in the Biological Sciences. Humana Press, pp. 135-150. ISBN 978-1-4612-6671-6

URL: http://link.springer.com/chapter/10.1007%2F978-1-4...
DOI: 10.1007/978-1-4612-0229-5_7

Abstract

Large-format 2D gel electrophoresis systems have been developed that are capable of resolving several thousand cellular proteins in a matter of days [1,2]. For a number of years, a combination of Edman microsequence analysis and identification of proteins by staining with specific antibodies has been used to systematically categorize proteins and establish cellular databases [3–5]. There are, however, significant problems associated with these approaches. Most resolved proteins are only present in the low- to upper-femtomole range, significantly below the level at which automated sequencers can reliably operate [6, 7]. The relatively slow speed of the Edman process (one or two samples per machine per day) also means that the sheer number of proteins is too great to permit large-scale characterization within any useful period of time. The use of monoclonal antibodies, whilst both rapid and extremely sensitive, requires the ready availability of a large pool of antibody probes.

Item Type: Book Section
Subjects: Investigative techniques and equipment > spectroscopy > mass spectrometry
CSHL Authors:
Communities: CSHL labs > Pappin lab
Depositing User: Matt Covey
Date: 1996
Date Deposited: 22 May 2014 19:43
Last Modified: 24 Feb 2017 17:10
URI: https://repository.cshl.edu/id/eprint/30142

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