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en:aree:proteomica:ms:start [2018/08/10 15:19] Gianluca Frustagli English translation. |
en:aree:proteomica:ms:start [2020/09/29 13:22] Gianluca Frustagli [Mass Spectrometry (MS)] Referent change. |
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| </WRAP> | </WRAP> | ||
| - | {{:aree:proteomica:ms:lab.jpeg?400 |}} The unit is equipped with an **LTQ XL Ion Trap** mass spectrometer from ThermoFisher with an additional ETD (Electron Transfer Dissociation) fragmentation(([[http://www.pnas.org/cgi/content/full/101/26/9528|Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry]])). | + | {{:aree:proteomica:ms:lab.jpeg?400 |}} The unit is equipped with two mass spectrometer from ThermoFisher, an **Orbitrap Fusion** and a **LTQ XL Ion Trap** with an additional ETD (Electron Transfer Dissociation) fragmentation(([[http://www.pnas.org/cgi/content/full/101/26/9528|Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry]])). |
| - | This instrument, in-line with a nano-capillary chromatograph, allows the separation of peptide mixtures with different complexity degree and the subsequent identification of the constituent proteins through dedicated software (LC-MS / MS analysis). This instrumentation is of great support in the biomedical research projects that require the identification and/or characterization of proteins and their level of expression. The samples to be analyzed can be multiple: synthetic proteins, semi-purified proteins, immuno-precipitation, sub-proteomes, total proteomes. Generally, the LC-MS / MS analysis is preceded by electrophoresis (PAGE) for a qualitative and quantitative estimate of the sample followed by protein in-gel digestion. | + | These instruments, in-line with nano-capillary chromatographs, allows the separation of peptide mixtures with different complexity degree and the subsequent identification of the constituent proteins through dedicated software (LC-MS / MS analysis). This instrumentation is of great support in the biomedical research projects that require the identification and/or characterization of proteins and their level of expression. The samples to be analyzed can be multiple: synthetic proteins, semi-purified proteins, immuno-precipitation, sub-proteomes, total proteomes. Generally, the LC-MS / MS analysis is preceded by electrophoresis (PAGE) for a qualitative and quantitative estimate of the sample followed by protein in-gel digestion. |
| In case of very poor samples or for methodological needs, the digestion can be carried out in solution and followed by off-line chromatography before the LC-MS /MS analysis(([[http://www.nature.com/nmeth/journal/v6/n5/abs/nmeth.1322.html|Universal sample preparation method for proteome analysis]])). In comparative studies, relative quantitative determination is possible through isotopic labeling techniques(([[https://www.ncbi.nlm.nih.gov/pubmed/17902639|iTRAQ Reagent-Based Quantitative Proteomic Analysis on a Linear Ion Trap Mass Spectrometer]]))(([[http://www.neurology.org/content/79/10/1012.abstract|Increased levels of acute-phase inflammatory proteins in plasma of patients with sporadic CJD]])) or label-free(([[https://www.ncbi.nlm.nih.gov/pubmed/23794183|Critical assessment of proteome-wide label-free absolute abundance estimation strategies]])). The unit relies particularly on extensive experience in the following areas: | In case of very poor samples or for methodological needs, the digestion can be carried out in solution and followed by off-line chromatography before the LC-MS /MS analysis(([[http://www.nature.com/nmeth/journal/v6/n5/abs/nmeth.1322.html|Universal sample preparation method for proteome analysis]])). In comparative studies, relative quantitative determination is possible through isotopic labeling techniques(([[https://www.ncbi.nlm.nih.gov/pubmed/17902639|iTRAQ Reagent-Based Quantitative Proteomic Analysis on a Linear Ion Trap Mass Spectrometer]]))(([[http://www.neurology.org/content/79/10/1012.abstract|Increased levels of acute-phase inflammatory proteins in plasma of patients with sporadic CJD]])) or label-free(([[https://www.ncbi.nlm.nih.gov/pubmed/23794183|Critical assessment of proteome-wide label-free absolute abundance estimation strategies]])). The unit relies particularly on extensive experience in the following areas: | ||
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| Several bioinformatic analysis approaches are available for the analysis of results. | Several bioinformatic analysis approaches are available for the analysis of results. | ||
| - | <wrap em>Service requests to this unit must be sent by filling out the '[[richiesta|Service Request]]' online form. You will be contacted for a preliminary meeting.</wrap> | + | \\ |
| + | |||
| + | === To ask for the service: === | ||
| + | |||
| + | - preliminary contact the [[chi_siamo|MS unit staff]]; | ||
| + | - fill out the online form '**[[richiesta|Service Request]]**' briefly illustrating the scientific problem, type and number of samples etc.; | ||
| + | - plan a meeting with the unit staff to discuss methods of analysis, costs, timing etc. | ||
| <wrap em>We recommend that you follow the advices in the '[[linee_guida|Guidelines]]' section during sample preparation.</wrap> | <wrap em>We recommend that you follow the advices in the '[[linee_guida|Guidelines]]' section during sample preparation.</wrap> | ||
| - | > Contact: [[marta.ponzi@iss.it|Marta Ponzi]] | + | \\ |
| - | > ☎ **+39 06 4990 3134** | + | |
| + | > Contact: [[serena.camerini@iss.it|Serena Camerini]] | ||
| + | > ☎ **+39 06 4990 3164** | ||
| + | > Fax **+39 06 4990 2040** | ||
| > Where we are: building **30**, floor **A**, room **11** | > Where we are: building **30**, floor **A**, room **11** | ||
| - | > FIXME **To be revised.**\\ //(remove this paragraph finished)// | + | /* > FIXME **To be revised.**\\ //(remove this paragraph finished)// */ |
