Dataset Information

Mouse_Hippocampus_LCMSMS_IEF_Pooling

ABSTRACT: Protein extract from a single mouse hippocampus was enzymatically digested and fractionated by isoelectric focusing. Aliquots of fractions were pooled to make fewer, more complex samples. The unfractionated lysate, fractions, and pooled fractions were subjected to liquid chromatographyâ??mass spectrometry analysis. Samples consisting of many individual fractions had more protein identifications and quantified proteins with more spectral counts and greater precision than protein extract that was unfractionated or pooled into fewer LC-MS/MS samples.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Richard Nowakowski

PROVIDER: MSV000084999 | MassIVE | Fri Feb 21 13:28:00 GMT 2020

SECONDARY ACCESSION(S): PXD002905

REPOSITORIES: MassIVE

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Publications

Fractionation-dependent improvements in proteome resolution in the mouse hippocampus by IEF LC-MS/MS.

Bundy Joseph L JL Inouye Brian D BD Mercer Roger S RS Nowakowski Richard S RS

Electrophoresis 20160601 14

An assessment of fractionated mouse hippocampal peptides was conducted. Protein extract from a single mouse hippocampus was enzymatically digested and fractionated by IEF. Aliquots of fractions were pooled into fewer, more complex samples. The unfractionated lysate, fractions, and pooled fractions were subjected to LC-MS/MS analysis. Samples consisting of many individual fractions had more protein identifications, greater protein sequence coverage, and quantified proteins with more spectral coun ...[more]

PMID: 27129779

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Project description:Part I - SI-NET tumors High Resolution Isoelectric Focusing (HiRIEF) LC-MS and relative quantification by iTRAQ 8-plex was used to analyze 14 small intestinal neuroendocrine tumors (SI-NETs). The data was obtained from two separate TMT10plex sets and linked by a single internal pooled standard. The internal pooled standard was made by combining equal aliquots of the tryptic peptide mixtures from each of the 14 tissue samples. iTRAQ set1 was composed of 7 SI-NET samples, all from different individuals, and internal pooled standard labelled as follows: Channel 113 (sample Screen-1 with liver metastasis), Channel 114 (sample Screen-8 with liver metastasis), Channel 115 (sample Screen-3 with liver metastasis), Channel 116 (sample Screen-2 with liver metastasis), Channel 117 (sample Screen-5 no liver metastasis), Channel 118 (sample Screen-4 no liver metastasis), Channel 119 (sample Screen-10 no liver metastasis), Channel 121 (internal pooled standard). iTRAQ set2 was composed of 7 SI-NET samples, all from different individuals, and internal pooled standard labelled as follows: Channel 113 (sample Screen-7 with liver metastasis), Channel 114 (sample Screen-11 with liver metastasis), Channel 115 (sample Screen-13 with liver metastasis), Channel 116 (sample Screen-6 no liver metastasis), Channel 117 (sample Screen-12 no liver metastasis), Channel 118 (sample Screen-9 no liver metastasis), Channel 119 (sample Screen-14 no liver metastasis), Channel 121 (internal pooled standard). Part II - time course profiling in cell lines High Resolution Isoelectric Focusing (HiRIEF) LC-MS and relative quantification by TMT 10-plex was used to analyze cellular response to the neddylation inhibitor pevonedistat (MLN4924) at different timepoints in two SI-NET (small intestinal neuroendocrine tumors) cell lines. The data was obtained from two separate TMT 10-plex experiments. TMT set1 includes a time course experiment upon pevonedistat treatment of CNDT2 cells with harvests at 2h, 6h, 12h and 24h after treatment as well as of untreated control cells. Isobaric tag labelling of peptide samples with TMT10plex was used as follows. Biological duplicate controls (TMT channels 126, 127N), duplicate 2h (127C, 128N), duplicate 6h (128C, 129N), duplicate 12h (129C, 130N) and duplicate 24h (130C, 131) samples were employed. TMT set2 includes a time course experiment upon pevonedistat treatment of HC45 cells with harvests at 2h, 6h, 12h and 24h after treatment as well as of untreated control cells. Isobaric tag labelling of peptide samples with TMT10plex was used as follows. Biological duplicate controls (TMT channels 126, 127N), duplicate 2h (127C, 128N), duplicate 6h (128C, 129N), duplicate 12h (129C, 130N) and duplicate 24h (130C, 131) samples were employed.

Dataset Information

Mouse_Hippocampus_LCMSMS_IEF_Pooling

Publications

Fractionation-dependent improvements in proteome resolution in the mouse hippocampus by IEF LC-MS/MS.

Similar Datasets

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