Journal
JOURNAL OF PROTEOMICS
Volume 73, Issue 3, Pages 571-578Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2009.09.003
Keywords
CD4/CD8 T-cells; Electrophoresis; MF10 fractionation; CD34 haematopoietic stem cells; Mass spectrometry; Proteome
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Funding
- Australian Research Council [LPO348904]
- Australian Postgraduate Award (Industry)
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Studying biological processes through protein expression is difficult due to the dynamic flux of the proteome, the extreme diversity and heterogeneity, the wide range of cellular protein expression, and the limited detection range of technology. Fractionating complex biological mixtures can help overcome these issues but it can be a challenging process particularly when fractionating rare samples (<= 10(6) cells), due to the absolute limits in protein copy number and abundance. In this study, partitioning by charge and mass using the Microflow MF10 has been applied to CD34(+) haematopoietic stem/progenitor cells and CD4(+)/CD8(+) T-cells to separate proteins restricted by cell number. Less than 10 mu g total proteins per fraction was used for comparative analysis using SDS-PAGE and identification of silver stained bands by LC-MS/MS revealed differentially expressed proteins between the 3 cell populations, which may be involved in cell differentiation. (C) 2009 Elsevier B.V. All rights reserved.
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