|
| Status |
Public on Mar 06, 2012 |
| Title |
preadipocytes, no differentiation, donor 3 |
| Sample type |
RNA |
| |
|
| Source name |
preadipocytes, no differentiation, donor 3
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: preadipocytes
time: 0 days of differentiation
individual: donor 3
|
| Treatment protocol |
Differentiation into adipocytes was initiated by addition of 10 µg/ml insulin, 1 µM dexamethasone, 200 µM indomethacin and 500 µM isobutylmethylxanthine (Cambrex) to the culture medium.
|
| Growth protocol |
Subcutaneous human preadipocytes were incubated in basal growth medium (Cambrex) containing 10% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicilline and 100 µg/ml streptomycine (= culture medium).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using Trizol (Invitrogen) following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
1 µg of each RNA was used as template to produce Cy3- labeled cRNA. The RNA samples were amplified and labeled using the Agilent Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Cy3 labeled cRNAs were hybridized overnight (17 hours, 65°C) to an Agilent Whole Mouse Genome Oligo Microarrays (44K) using Agilent’s recommended hybridization chamber and oven.Finally, microarrays were washed once with wash buffer 1 for 1 min at RT followed by a second wash with wash buffer 2 for 1 min at 37°C and a final wash step with acetonitrile for 30 sec at RT.
|
| Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent’s Microarray Scanner System (G2505C, Agilent Technologies, Palo Alto, USA).
|
| Description |
female, 34 y, BMI 22.24, subcutaneous abdomen
|
| Data processing |
The Agilent Feature Extraction Software (FES 10.5.1.1 ) was used to read out and process the microarray image files.
|
| |
|
| Submission date |
Apr 14, 2011 |
| Last update date |
Mar 06, 2012 |
| Contact name |
Silvia Rueberg |
| Organization name |
Miltenyi Biotec GmbH
|
| Department |
Genomic Services Department
|
| Street address |
Friedrich-Ebert-Str. 68
|
| City |
Bergisch Gladbach |
| ZIP/Postal code |
51429 |
| Country |
Germany |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE28628 |
Identification of new genes involved in human adipogenesis and fat storage |
|
