|
| Status |
Public on Nov 30, 2016 |
| Title |
Iso (1) |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
HEK293_agonist treated
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HEK293
treated with: 10nM isoproterenol for 2 hrs
|
| Treatment protocol |
Agonist added at indicated concentration for 2 hrs directly to medium
|
| Growth protocol |
HEK293 cells were grown in a CO2 incubator at 37C in DMEM+FBS medium; drug was added directly to medium for 2 hrs
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Kit
|
| Label |
Cy5
|
| Label protocol |
Amersham Cy dye kit
|
| |
|
| Channel 2 |
| Source name |
Untreated cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HEK293
treated with: none (untreated)
sample type: Reference
|
| Treatment protocol |
Agonist added at indicated concentration for 2 hrs directly to medium
|
| Growth protocol |
HEK293 cells were grown in a CO2 incubator at 37C in DMEM+FBS medium
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Kit
|
| Label |
Cy3
|
| Label protocol |
Amersham Cy dye kit
|
| |
|
| |
| Hybridization protocol |
Cy5- and Cy3-labeled aRNA were combined in 3× SSC, 50 mM Hepes (pH 70), 20 µg of poly(A) RNA, 20 µg Cot-1 DNA, 20 µg tRNA, and 03% SDS to a total volume of 50uL Samples were then heated to 70°C for 5 min, spun at 14,000rpm at room temperature in a microfuge for 5 min, then hybridized at 65°C using the MAUI hybridization system (BioMicro) for 12–16 hrs
|
| Scan protocol |
Microarrays were scanned using AxonScanner 4000B (Molecular Devices), and PMTs were manually adjusted for every slide scanned to maximize signal without saturation
|
| Description |
Sample 1
|
| Data processing |
GenePix was used to extract data from scanned microarray images Log2(Treated/Untreated) from duplicate sports were averaged and all log2 ratios were median-centered on an array-by-array basis
|
| |
|
| Submission date |
Sep 28, 2016 |
| Last update date |
Nov 30, 2016 |
| Contact name |
Nikoleta Tsvetanova |
| E-mail(s) |
nitsvetanova@gmail.com
|
| Organization name |
University of California, San Francisco
|
| Street address |
600 16th street
|
| City |
San Francisco |
| State/province |
CA |
| ZIP/Postal code |
94158 |
| Country |
USA |
| |
|
| Platform ID |
GPL11179 |
| Series (1) |
| GSE87461 |
GPCR endocytosis confers uniformity in responses to chemically distinct ligands |
|
