|
| Status |
Public on Feb 13, 2024 |
| Title |
Donor 1; Sivelestat_High; 10 min |
| Sample type |
protein |
| |
|
| Source name |
Human islets
|
| Organism |
Homo sapiens |
| Characteristics |
diabetes status: Non-diabetic
unos id/rrid: SAMN09768368
date of treatment: 8.8.18
|
| Treatment protocol |
Human islets were starved in Final Wash media for 3 hours. After, IEQs were treated with DMSO, telaprevir (1 and 100 µg/ml), tebipenem (1 and 100 µg/ml), sivelestat (100 µg/ml), 5-IT (1 µM), Ovalbumin (1 µg/ml) and SerpinB1 (1 µg/ml) for 10 or 30 minutes. At the end of the incubation, islets were collected and washed once in PBS. Finally islet pellet was snap-frozen in liquid nitrogen.
|
| Growth protocol |
Human islets were obtained by Prodo Lab or IIDP. Upon receipt, human islets were cultured in Miami Media at 37 C 5% CO2. After overnight incubation, 150/200 size-matched IEQs per condition were hand-picked for the following procedure
|
| Extracted molecule |
protein |
| Extraction protocol |
Total proteins were harvested from islets lysates using M-PER protein extraction reagent (Thermo Fisher, USA) supplemented with proteinase and phosphatase inhibitors (Sigma, USA) according to standard protocol.
|
| Label |
biotin
|
| Label protocol |
Proteins were covalently labeled with biotin. Free biotin molecules are then removed at the completion of labeling reactions by gel filtration.
|
| |
|
| Hybridization protocol |
After blocking non-specific binding sites on the array, an incubation chamber is mounted onto the microarray to permit the loading of 2 samples side by side on the same chip and prevent mixing of the samples. Following sample incubation, unbound proteins are washed away and the array is then probed with anti-biotin antibody that is labelled with a proprietary fluorescent dye combination.
|
| Scan protocol |
Each array produces a pair of 16-bit images, which are captured with a Perkin-Elmer ScanArray Reader laser array scanner (Waltham, MA).
|
| Description |
ID19924
|
| Data processing |
Signal quantification is performed with ImaGene 9.0 from BioDiscovery (El Segundo, CA) with predetermined settings for spot segmentation and background correction. The background-corrected raw intensity data are logarithmically transformed with base 2.
|
| |
|
| Submission date |
Dec 02, 2021 |
| Last update date |
Feb 13, 2024 |
| Contact name |
Jonathan M Dreyfuss |
| Organization name |
Joslin Diabetes Center
|
| Department |
Bioinformatics & Biostatistics Core
|
| Street address |
1 Joslin Pl
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02215 |
| Country |
USA |
| |
|
| Platform ID |
GPL26730 |
| Series (1) |
| GSE189986 |
Short-term effects of Elastase inhibition on human islets protein phosphorylation |
|
