Dataset Information

Miniaturized sample preparation on a digital microfluidics device for sensitive bottom-up microproteomics of mammalian cells using magnetic beads and mass spectrometry-compatible surfactants

ABSTRACT: We used digital microfluidics (DMF) for the sample preparation of approximately 100 mammalian cells, and subsequent bottom-up proteome analysis by LC-MS. This comprised optimization of cell lysis conditions for DMF, the development of detergent-buffer systems, and adaptation of the single-pot, solid-phase-enhanced sample preparation (SP3) approach on-chip. Application of the methodology to the proteome analysis of Jurkat T cells led to the identification of up to 2,500 proteins from approximately 500 cells, and up to 1,200 proteins from approximately 100 cells.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture, T Cell

SUBMITTER: Andreas Tholey

LAB HEAD: Andreas Tholey

PROVIDER: PXD014158 | Pride | 2019-10-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	02042019_BLANK_Urea-SP3-BBS.pep.pep.xml	Pepxml
	02042019_BLANK_Urea-SP3-BBS.prot.prot.xml	Xml
	02042019_BLANK_Urea-SP3-BBS.raw	Raw
	07052019_15kJurkats_10u_190511030118.pep.pep.xml	Pepxml
	07052019_15kJurkats_10u_190511030118.prot.prot.xml	Xml

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Miniaturized sample preparation on a digital microfluidics device for sensitive bottom-up microproteomics of mammalian cells using magnetic beads and mass spectrometry-compatible surfactants.

Leipert Jan J Tholey Andreas A

Lab on a chip 20191001 20

While LC-MS-based proteomics with high nanograms to micrograms of total protein has become routine, the analysis of samples derived from low cell numbers is challenged by factors such as sample losses, or difficulties encountered with the manual manipulation of small liquid volumes. Digital microfluidics (DMF) is an emerging technique for miniaturized and automated droplet manipulation, which has been proposed as a promising tool for proteomic sample preparation. However, proteome analysis of sa ...[more]

PMID: 31531506

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