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MB Sample ID: SA028919
Local Sample ID: | IgAN1_4 |
Subject ID: | SU000582 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Cell Strain Details: | Suzuki, H., Moldoveanu, Z., Hall, S., et al. IgA1-secreting cell lines from patients with IgA nephropathy produce aberrantly glycosylated IgA1. J Clin Inv. 2008, 118, 629-639 |
Cell Primary Immortalized: | immortalized |
Cell Counts: | 1x10^7 cells / pellet |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000582 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Cell Strain Details: | Suzuki, H., Moldoveanu, Z., Hall, S., et al. IgA1-secreting cell lines from patients with IgA nephropathy produce aberrantly glycosylated IgA1. J Clin Inv. 2008, 118, 629-639 |
Cell Primary Immortalized: | immortalized |
Cell Counts: | 1x10^7 cells / pellet |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
IgAN1_4 | SA028919 | FL007042 | IgAN | Phenotype |
Collection:
Collection ID: | CO000576 |
Collection Summary: | An aliquot of cells (1 x 10^7) grown in suspension culture was transferred into a 15 mL conical tube containing 4x the culture volume of ice-cold 0.9% (w/v) NaCl. Cells were pelleted for 5 minutes at 1,500 rpm in a 4oC refrigerated centrifuge. Supernatant was removed, and the pellet was gently resuspended in 1 mL of ice-cold 0.9% (w/v) NaCl to wash and transfer into a 2 mL Lo-Bind microcentrifuge tube (Eppendorf, #022431102). Cells were pelleted for 3 minutes at 1,300 rcf in a 4oC refrigerated centrifuge. The entire supernatant was removed and pellets were snap-frozen in liquid nitrogen and immediately stored at -70°C. |
Sample Type: | Cell pellets |
Storage Conditions: | -80 C |
Tissue Cell Quantity Taken: | 1 x 10^7 |
Treatment:
Treatment ID: | TR000596 |
Treatment Summary: | None |
Cell Media: | 10%FBS, 11mM Glucose |
Sample Preparation:
Sampleprep ID: | SP000589 |
Sampleprep Summary: | A total of 24 cell pellets (4 replicates from 6 cell lines) were shipped to the NIH RTI-RCMRC on dry ice and immediately stored at -80oC after being inventoried for metabolomics analysis. An addition of 500 µL of ice-cold Cell Extraction buffer (0.005 tryptophan-d5 in 50:50 Acetonitre:Water) was added to tubes containing the cell pellet samples on ice. MagNA Lyser ceramic beads (~15-20, prewashed & dried) were added to the tubes and the MagNA Lyser was used to beat samples for two 30 sec pulses at 2,000 rpm and samples were placed on a cold block for 1 min between pulses. Samples were then centrifuged at room temperature at 16,000 rcf for 4 min. A 15 mL washed conical tube was labeled Total Pool and 120 µL of each sample was added to this conical tube. There were 24 samples; thus, the resulting Total Pool was 2880 µL. The Total Pool was vortexed and 150 µL of the Total Pool was transferred to pre-labeled 1.5 mL Lo-Bind Eppendorf tubes to make two sets of 5 Total Pool samples, and 3 Equilibrium samples. The 150 uL of the remaining supernatants of each sample was transferred to a new, pre-labeled 1.5 mL Lo-Bind Eppendorf tubes and capped with disposable rubber stoppers. Samples were placed at -80 °C for 1 h and lyophilized overnight. 100 µL of 95:5 Acetonitrile:Water was added to each tube and vortexed for 2 min at 5000 rpm, followed by centrifugation at room temperature at 16,000 rcf for 4 min. The supernatants were transferred to pre-labeled autosampler vials and 3 µL was injected into SYNAPT G2-Si. All samples were prepared and analyzed in a randomized order. |
Combined analysis:
Analysis ID | AN000860 | AN000861 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | HILIC |
Chromatography system | Waters Acquity I-Class | Waters Acquity I-Class |
Column | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | QTOF | QTOF |
MS instrument name | Waters Synapt G2 Si QTOF | Waters Synapt G2 Si QTOF |
Ion Mode | POSITIVE | NEGATIVE |
Units | normalized ion counts | normalized ion counts |
Chromatography:
Chromatography ID: | CH000612 |
Chromatography Summary: | Hydrophilic Interaction Liquid Chromatography (HILIC) Gradient Seperation |
Instrument Name: | Waters Acquity I-Class |
Column Name: | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
Chromatography Type: | HILIC |
MS:
MS ID: | MS000761 |
Analysis ID: | AN000860 |
Instrument Name: | Waters Synapt G2 Si QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |
MS ID: | MS000762 |
Analysis ID: | AN000861 |
Instrument Name: | Waters Synapt G2 Si QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |