PMID:16461942
| Citation |
Shi, Y, Hill, J, Xiang, R, Horváth, C and Wilkins, JA (2005) Multistep mass tagging coupled with 2D LC-MS--an approach to increasing the number of identified proteins. J Biomol Tech 16:190-6 |
|---|---|
| Abstract |
Identification of large numbers of proteins from complex biological samples is a continuing challenge in the area of quantitative proteomics. We introduce here a simple and reliable multistep mass tagging technique using our recently developed solid phase mass tagging reagents. When coupled with two-dimensional liquid chromatography/nano-electrospray ionization ion trap mass spectrometry (2D-LC/nano-ESI-MS), this method allows enhanced protein identification when tested on samples from prokaryotic and eukaryotic sources. The proteome of Escherichia coli D21 grown to either mid-exponential or stationary phase, and the membrane proteome from established breast cancer cell lines BT474 and MCF7 were used as model systems in these experiments. In both experiments, the numbers of total identified proteins are at least twice the numbers identified from a single tagging cycle. The sample complexity can be effectively reduced with corresponding increases in protein identification using the multistep method. The strategy described here represents a potentially powerful technique for large-scale qualitative and quantitative proteome research. |
| Links | |
| Keywords |
Cell Line, Tumor; Chromatography, Liquid/methods; Escherichia coli Proteins/chemistry; Humans; Neoplasm Proteins/chemistry; Proteome; Spectrometry, Mass, Electrospray Ionization/methods |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.