Summary of Study ST001333
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000910. The data can be accessed directly via it's Project DOI: 10.21228/M82H5K This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST001333 |
| Study Title | Huntington’s Disease Genotype Suppresses Global Manganese-Responsive Processes |
| Study Type | Untargeted metabolomics analysis |
| Study Summary | Global untargeted metabolomics studies performed in the striatum tissue, the brain region most sensitive to neurodegeneration in Huntington’s Disease, to investigate global Mn-dependent and Mn-responsive biology following various Mn exposures in a mouse model of HD. |
| Institute | Vanderbilt University |
| Department | Chemistry |
| Laboratory | Center for Innovative Technology |
| Last Name | Codreanu |
| First Name | Simona |
| Address | 1234 Stevenson Center Lane |
| simona.codreanu@vanderbilt.edu | |
| Phone | 6158758422 |
| Submit Date | 2020-03-27 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2020-09-29 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Sample Preparation:
| Sampleprep ID: | SP001415 |
| Sampleprep Summary: | Previously frozen striatal tissue was lysed in 400µL ice-cold lysing buffer (1:1:2, ACN:MeOH:Ammonium Bicarbonate (0.1M, pH 8.0) (LC-MS grade). Individual samples were sonicated using a probe tip sonicator, 10 pulses, at 30% power and cooled down on ice between samples. A BCA protein assay was used to determine the protein concentration for each individual sample, and adjusted to a total amount of protein of 200µg in 200 µL of lysis buffer. Isotopically labeled standards, Creatinine-D3 and Lysine-D4, were added to each sample to assess sample processing steps (metabolite extraction and reconstitution). Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum. Dried metabolite extracts were stored frozen at -80C until ready to use. Prior to mass spectrometry analysis, extracts were reconstituted in 100 μl of acetonitrile/ water (80:20, v/v) and centrifuged for 5 min at 15K rpm to remove insoluble material. Quality control samples were prepared by pooling equal volumes of each sample. Isotopically labeled standards, Valine-D8 and Inosine-4N15, were added to each sample to determine MS instrument reproducibility. |
| Sampleprep Protocol Filename: | Codreags00_20200327_075634_PR_MS_Metabolomics_Protocol.pdf |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum. |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 100 μl of acetonitrile/ water (80:20, v/v) and centrifuged for 5 min at 15K rpm to remove insoluble material |
| Sample Spiking: | Valine-D8 and Inosine-4N15, were added to each sample to determine MS instrument reproducibility. |
