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MB Sample ID: SA273747

Local Sample ID:04_B0_Spleen_syngenic_42days_170427_UKy_GCH_rep1-polar-ICMS_A
Subject ID:SU002825
Subject Type:Mouse
Subject Species:Mus musculus
Taxonomy ID:10090

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Subject:

Subject ID:SU002825
Subject Type:Mouse
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
04_B0_Spleen_syngenic_42days_170427_UKy_GCH_rep1-polar-ICMS_ASA273747FL035486syngenicTreatment
04_B0_Spleen_syngenic_42days_170427_UKy_GCH_rep1-polar-ICMS_ASA273747FL03548642Time Point

Collection:

Collection ID:CO002818
Collection Summary:Mouse is sacrificed and tissues are harvested.
Collection Protocol ID:mouse_tissue_collection
Collection Protocol Filename:mouse_tissue_procedure.pdf
Sample Type:Multiple tissues

Treatment:

Treatment ID:TR002834
Treatment Summary:Mouse with allogenic bone marrow transplant. Fed with semi-liquid diet supplemented with fully labeled glucose for 24 hours before harvest.;Mouse with no treatment. Fed with semi-liquid diet supplemented with fully labeled glucose for 24 hours before harvest.;Mouse with syngenic bone marrow transplant. Fed with semi-liquid diet supplemented with fully labeled glucose for 24 hours before harvest.
Treatment Protocol ID:allogenic;naive;syngenic
Treatment Protocol Filename:study_treatments.pdf

Sample Preparation:

Sampleprep ID:SP002831
Sampleprep Summary:Tissue is frozen in liquid nitrogen to stop metabolic processes.;Frozen tissue is ground in a SPEX grinder under liquid nitrogen to homogenize the sample.;Polar extraction from homogenate, lypholized, and frozen.;Protein extraction and quantification.;Before going into the IC-FTMS the frozen sample is reconstituted in water.
Sampleprep Protocol ID:tissue_quench;frozen_tissue_grind;polar_extraction;protein_extraction;IC-FTMS_preparation
Sampleprep Protocol Filename:No tissue_quench file.
No frozen_tissue_grind file.
4B_Extract_Polar_Lipid_Prot_Fan_070417.pdf
['4D_17Jun4_Fan_Prot_Quant.pdf', '4B_Extract_Polar_Lipid_Prot_Fan_070417.pdf']
No IC-FTMS_preparation file.

Combined analysis:

Analysis ID AN004409
Analysis type MS
Chromatography type Targeted IC
Chromatography system Thermo Dionex ICS-5000+
Column Dionex IonPac AS11-HC-4um 2 mm i.d. x 250 mm
MS Type ESI
MS instrument type IC-FTMS
MS instrument name Orbitrap Fusion
Ion Mode NEGATIVE
Units natural abundance corrected and dry residue normalized peak area

Chromatography:

Chromatography ID:CH003309
Chromatography Summary:Targeted IC
Instrument Name:Thermo Dionex ICS-5000+
Column Name:Dionex IonPac AS11-HC-4um 2 mm i.d. x 250 mm
Column Temperature:--
Flow Gradient:--
Flow Rate:--
Solvent A:--
Solvent B:--
Chromatography Type:Targeted IC

MS:

MS ID:MS004157
Analysis ID:AN004409
Instrument Name:Orbitrap Fusion
Instrument Type:IC-FTMS
MS Type:ESI
MS Comments:ICMS Analytical Experiment with detection of compounds by comparison to standards. Thermo RAW files are loaded into TraceFinder and peaks are manually curated. The area under the chromatograms is then exported to an Excel file. The area is then corrected for natural abundance. The natural abundance corrected area is then used to calculate the concentration of each compound for each sample. This calculation is done using standards. The first sample ran on the ICMS is a standard that has known concentrations of certain compounds. Then a number of samples are ran (typically 3-4) followed by another standard. The equation to calculate the concentration is "intensity in sample"/("intensity in first standard" + (("intensity in second standard" - "intensity in first standard")/# of samples) * "known concentration in standard", where the "intensity" is the aforementioned natural abundance corrected area, and the unlabeled intensity from the standard is used for all isotopologues of the compound. The reconstitution volume is simply the volume that the polar part of the sample was reconstituted to before going into the ICMS. The injection volume is how much of the reconstitution volume was injected into the ICMS. The protein is how much protein was in the entire sample (not only the small portion that was aliquoted for the ICMS). The polar split ratio is the fraction of the polar part of the sample that was aliquoted for the ICMS. This is calculated by dividing the weight of the polar aliquot for ICMS by the total weight of the polar portion of the sample. The protein normalized concentration is calculated using the equation, concentration * (reconstitution volume / 1000 / polar split ratio / protein).
Ion Mode:NEGATIVE
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