|
| Status |
Public on Jul 23, 2010 |
| Title |
Magnetically sorted untouched CD8+CD45R0-T cells mRNA from a three donors pooled sample unstimulated |
| Sample type |
RNA |
| |
|
| Source name |
Human CD8 T cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD8+CD45R0-T cells
stimulated: no
donor pool ethnicity: Caucasian
health status: healthy
|
| Treatment protocol |
IFN alfa 2 or IFN alfa 5 at 500 IU/mL
|
| Growth protocol |
Untouched CD8+CD45RO-cells were cultured at a concentration of 500.000 cells/mL with culture medium alone (resting) or culture medium containing IFNa2 or IFNa5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The cells were harvested with TRIzol Reagent (Invitrogen) and the RNA was extracted according to the manufacturer's instructions. As a last step of the extraction procedure, the RNA was purified with the RNeasy Mini-kit (Qiagen, Hilden, Germany).
|
| Label |
biotin
|
| Label protocol |
The cDNA was prepared from 5 microg total RNA using the One –Cycle cDNA Synthesis kit from Affymetrix. This cDNA was used for synthezising the cRNA using the IVT labeling kit (Affimetrix). The synthezised cRNA was purified with the GeneChip Sample Cleanup Module (Affimetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 microg of cRNA were hybridized to the Affymetrix HG-U133A 2.0 chip according to the protocols provided by the manufacturer.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 from Affymetrix.
|
| Description |
three donors pooled sample
|
| Data processing |
RMA
|
| |
|
| Submission date |
Jul 24, 2009 |
| Last update date |
Jul 24, 2009 |
| Contact name |
Iranzu Gonzalez |
| URL |
http://www.cima.es
|
| Organization name |
Center for applied medical research
|
| Department |
Hepathology and Gene Therapy
|
| Lab |
3.03. Clinic immunology
|
| Street address |
Avda. Pio XII nº 55
|
| City |
Pamplona |
| State/province |
Navarra |
| ZIP/Postal code |
31008 |
| Country |
Spain |
| |
|
| Platform ID |
GPL571 |
| Series (2) |
| GSE17299 |
Direct effects of IFNα on human CD8 T cells_without any other concomitant signals |
| GSE17302 |
IFNα-mediated gene expression patterns |
|
