|
| Status |
Public on Nov 07, 2008 |
| Title |
RL120502-mono macro |
| Sample type |
RNA |
| |
|
| Source name |
brain; treated with cytokines typical of monocyte/macrophages
|
| Organism |
Rattus norvegicus |
| Characteristics |
CNS mixed glial cultures from 2-3 day old Sprague-Dawley rats
|
| Treatment protocol |
Cells were maintained in defined medium with 2% fetal bovine serum for 6-8 days, then treated with cytokine mixtures representative of Th1, Th2 or monocyte/macrophage cytokines for 6 hours (Lisak, et al., Mult. Scler. 12:149-168, 2006)
|
| Growth protocol |
Mixed CNS glial cultures were prepared from neonatal rat brain using a modification of the shakeoff technique (McCarthy and deVellis, J. Cell. Biol. 85:890-902, 1980; Dyer and Benjamins, J. Neurosci. 8:4307-4318, 1988)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cells 2-3 times for each time point. Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 20 ug of cRNA were hybridized for 16 hr at 45C on Rat RG-U34A Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Agilent GeneArray Scanner. Normalization Factor- 1.0, TGT=500 (all probe sets)
|
| Description |
Treated with cytokines typical of monocyte/macrophages for 6 hours; recombinant rat IL1a, IL1b, IL-12p40, and TNF-a, all at 10 ng/ml in defined medium with 2% fetal bovine serum.
|
| Data processing |
All cel files were read and normalized by dChip v1.2, normalized data were analysed by GeneSpring software
|
| |
|
| Submission date |
Nov 21, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
wenbo xu |
| Organization name |
wayne state university
|
| Street address |
3127 Scott Hall ,540 East Canfield
|
| City |
Detroit |
| State/province |
MI |
| ZIP/Postal code |
48201 |
| Country |
USA |
| |
|
| Platform ID |
GPL85 |
| Series (1) |
| GSE9659 |
Expression data for rat CNS mixed glial cultures treated with cytokines |
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