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Citation
Hughes, C.S., Foehr, S., Garfield, D.A., Furlong, E.E., Steinmetz, L.M., Krijgsveld, J. (2014). Ultrasensitive proteome analysis using paramagnetic bead technology.  Mol. Syst. Biol. 10(10): 757.
FlyBase ID
FBrf0226641
Publication Type
Research paper
Abstract

In order to obtain a systems-level understanding of a complex biological system, detailed proteome information is essential. Despite great progress in proteomics technologies, thorough interrogation of the proteome from quantity-limited biological samples is hampered by inefficiencies during processing. To address these challenges, here we introduce a novel protocol using paramagnetic beads, termed Single-Pot Solid-Phase-enhanced Sample Preparation (SP3). SP3 provides a rapid and unbiased means of proteomic sample preparation in a single tube that facilitates ultrasensitive analysis by outperforming existing protocols in terms of efficiency, scalability, speed, throughput, and flexibility. To illustrate these benefits, characterization of 1,000 HeLa cells and single Drosophila embryos is used to establish that SP3 provides an enhanced platform for profiling proteomes derived from sub-microgram amounts of material. These data present a first view of developmental stage-specific proteome dynamics in Drosophila at a single-embryo resolution, permitting characterization of inter-individual expression variation. Together, the findings of this work position SP3 as a superior protocol that facilitates exciting new directions in multiple areas of proteomics ranging from developmental biology to clinical applications.

PubMed ID
PubMed Central ID
PMC4299378 (PMC) (EuropePMC)
Related Publication(s)
Note

Efficient sample processing for proteomics applications--are we there yet?
Kanshin and Thibault, 2014, Mol. Syst. Biol. 10: 758 [FBrf0227544]

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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Mol. Syst. Biol.
    Title
    Molecular Systems Biology
    Publication Year
    2005-
    ISBN/ISSN
    1744-4292
    Data From Reference