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MB Sample ID: SA035098
Local Sample ID: | NTM20L1 |
Subject ID: | SU000647 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000647 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
NTM20L1 | SA035098 | FL007497 | Control | Condition |
Collection:
Collection ID: | CO000641 |
Collection Summary: | Donor tissues were used following the institutional review board approved protocols and adhering to the tenets of the Declaration of Helsinki. Control TM and a subset of primary open-angle glaucoma (POAG) TM were collected from cadaver donors, a limited subset of POAG surgical specimen (acquired under institutional review board approved/exempted protocols) also were used for these studies. Cadaver donor tissues were sourced from Midwest Eye-Banks, Lions Eye Bank (Miami, FL), National Disease Research Interchange (NDRI; Philadelphia, PA), and Mundorf Eye Institute (Charlotte, NC). The postmortem time ranged from 3 to 16 hours. Control TM was stored briefly in Optisol at 48 and then used for lipid extraction. POAG TM tissues were stored at -80˚C until time of use. The mean age of control donors was 58.2 +/- 13.7 years and the mean age of glaucomatous TM donors 66 +/- 9.5 years. |
Sample Type: | Eye tissue |
Treatment:
Treatment ID: | TR000661 |
Treatment Summary: | No treament was performed on tissue. |
Sample Preparation:
Sampleprep ID: | SP000654 |
Sampleprep Summary: | Isolated trabecular meshwork tissue was subjected to an alternating cycle of immersion in liquid nitrogen and 37˚C, followed by extraction of lipids using a modified Bligh and Dryer method.13 The organic phase with extracted lipids was dried in a Speed-Vac (Model 7810014; Labconco, Kansas City, MO). Samples were flushed with argon gas to prevent oxidation. Corresponding aqueous phase extracted proteins were subjected to determination of concentration using Bradford’s method. A subset of protein samples also were subjected to densitometric quantification using bovine serum albumin as a standard (amino acid quantified) after electrophoretic separation on a PHAST (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) gel system. All extractions and subsequent handling were made using glass vials, and polyvinyl plastic was avoided completely. We also added a PC control standard during tissue homogenization, determined its recovery in an aliquot, and used for the calculation of total recovery of this standard for each extraction to ensure >99% recovery of added standard during extraction. |
Combined analysis:
Analysis ID | AN000956 |
---|---|
Analysis type | MS |
Chromatography type | None (Direct infusion) |
Chromatography system | None |
Column | none |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | Thermo Quantiva Access Max |
Ion Mode | UNSPECIFIED |
Units | Peak area |
Chromatography:
Chromatography ID: | CH000681 |
Instrument Name: | None |
Column Name: | none |
Chromatography Type: | None (Direct infusion) |
MS:
MS ID: | MS000851 |
Analysis ID: | AN000956 |
Instrument Name: | Thermo Quantiva Access Max |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | UNSPECIFIED |