MassIVE MSV000080744

Imported Reanalysis Dataset Public PXD005402

Use of a modified Multiplexed Inhibitor Bead assay to study kinome responses to cytostatic treatment in human cancer cell lines Triplicate Test: pilot study

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Description

In order to improve elution of proteins from the column, to reduce the cost as well as the hands on time of the analysis, we here present a modification of the Multiplexed Inhibitor Bead assay (MIB assay), where we elute the proteins from beads containing covalently bound inhibitors using trypsin digestion. Three human cancer cell lines treated with the same chemotherapeutics, cisplatin and docetaxel, were used to monitor and evaluate the performance of the assay. Using this method, we see a good coverage of the kinome as well as other important signaling proteins using only three kinase inhibitors. We demonstrate that the method is reproducible and that the column is re-usable for more than 10 times without loss in performance. The MIB assay revealed large differences in the cellular signaling responses between the cell lines. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: MS ; test ; triplicate

Contact

Principal Investigators:
(in alphabetical order) 		Voin Petrovic, Department of Cancer Research and Molecular Medicine Department of Language and Literature Erling Skjalgssons g 1, aboratoriesenteret * * Laboratoriesenter Norwegian University of Science and Technology Trondheim 7030 Norway, N/A
Submitting User: ccms

Publications

Petrovic V, Olaisen C, Sharma A, Nepal A, Bugge S, Sundby E, Hoff BH, Slupphaug G, Otterlei M.
On-column trypsinization allows for re-use of matrix in modified multiplexed inhibitor beads assay.
Anal. Biochem. Epub 2017 Mar 3.

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Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

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Number of distinct proteins found to be differentially abundant in at least one comparison across all analyses (original submission and reanalyses) associated with this dataset.

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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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This dataset may not contain all raw spectra data as originally deposited in PRIDE. It has been imported to MassIVE for reanalysis purposes, so its spectra data here may consist solely of processed peak lists suitable for reanalysis with most software.