Summary of Study ST002445

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 PR001576. The data can be accessed directly via it's Project DOI: 10.21228/M80709 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	ST002445
Study Title	The interaction of ceramide-1-phosphate with group IVA cytosolic phospholipase A2 modulates neutrophil polarization during inflammatory responses
Study Summary	Bone marrow-derived mouse neutrophils from wildtype, cPLA2alpha-knockin (interaction site on cPLA2alpha for its substrate C1P was ablated), and cPLA2alpha-knockout (cPLA2alpha gene mutated) mice were exposed to 4 hours of trans-endothelial migration and resulting eicosanoids were analyzed (eg, PGE2, PGD2, 5-HETE, and 5-oxo-ETE).
Institute	University of South Florida
Last Name	Maus
First Name	Kenneth
Address	4202 E Fowler Ave, CMMB - NES 107 - Chalfant Lab
Email	kmaus@usf.edu
Phone	8139283137
Submit Date	2023-01-16
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2023-02-05
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002540
Sampleprep Summary:	Media collection: Add proper volumes to achieve final concentrations of 10% MeOH and 0.5% glacial acetic acid. Then add 20 μL eicosanoid internal standard. Example: For 4 mL media add 400 μL of 100% MeOH and 20 μL of glacial acetic acid. • Example, for 24 samples, prep for 26: o 2.6 mL 100% MeOH (100 μL x 26) o 130 μL glacial acetic acid (5 μL x 26) o 520 μL eicosanoid IS (20 μL x 26) Columns: Set the manometer on the vacuum chamber to 5 mmHg. Precondition columns: Elute 2 mL MeOH, stop, then elute 2 mL H2O. Load sample and tighten the caps. Wash: Add 2 mL of 5% MeOH to the sample vial. Vortex and apply to the column under vacuum. Allow to run dry for 30 seconds. Stop vacuum, remove manifold & lay on side. Change out glass vials and elute: Apply 2 mL Isopropyl alcohol to column and equilibrate for 1 minute. Elute under vacuum and run dry for 30 seconds. Concentration: The solvent is removed by speed vac for !5 hours. Eicosanoids are re-dissolved in 100 μL of 50% EtOH in HPLC water. Centrifuge @ 4000 RPM for 10 minutes to pellet. Transfer supernatant into glass 0.1 mL mini conicals and load into white slots in HPLC tray.

Sampleprep ID:

SP002540

Sampleprep Summary:

Media collection: Add proper volumes to achieve final concentrations of 10% MeOH and 0.5% glacial acetic acid. Then add 20 μL eicosanoid internal standard. Example: For 4 mL media add 400 μL of 100% MeOH and 20 μL of glacial acetic acid. • Example, for 24 samples, prep for 26: o 2.6 mL 100% MeOH (100 μL x 26) o 130 μL glacial acetic acid (5 μL x 26) o 520 μL eicosanoid IS (20 μL x 26) Columns: Set the manometer on the vacuum chamber to 5 mmHg. Precondition columns: Elute 2 mL MeOH, stop, then elute 2 mL H2O. Load sample and tighten the caps. Wash: Add 2 mL of 5% MeOH to the sample vial. Vortex and apply to the column under vacuum. Allow to run dry for 30 seconds. Stop vacuum, remove manifold & lay on side. Change out glass vials and elute: Apply 2 mL Isopropyl alcohol to column and equilibrate for 1 minute. Elute under vacuum and run dry for 30 seconds. Concentration: The solvent is removed by speed vac for !5 hours. Eicosanoids are re-dissolved in 100 μL of 50% EtOH in HPLC water. Centrifuge @ 4000 RPM for 10 minutes to pellet. Transfer supernatant into glass 0.1 mL mini conicals and load into white slots in HPLC tray.