|
| Status |
Public on Oct 16, 2009 |
| Title |
Patient7C |
| Sample type |
RNA |
| |
|
| Source name |
Unchallenged CD4+ cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD4+ cells
condition: Unchallenged
subject: Patient7
|
| Treatment protocol |
PBMCs were cultured with diluent (PBS) or diluent with 100 ug grass pollen extract (ALK-Abelló A/S, Hörsholm, Denmark).
|
| Growth protocol |
PBMCs were prepared by centrifugation on Ficoll-Hypaque, washed and stimulated in microtiter wells with grass pollen extract (ALK-Abelló A/S, Hörsholm, Denmark; 100 ug/mL) and cell culture medium at a density of 106 cells/mL in a total volume of 2 mL per well. After seven days of incubation at 37 oC and 5 % CO2 the CD4+ cells were enriched from the allergen-challenged PBMCs by using anti-CD4-coated paramagnetic microbeads and a MACS system according to the instructions of the manufacturer (Miltenyi Biotech GmbH, Bergisch Gladbach, Germany).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
200 ug of total RNA was prepared using Ambion's Illumina RNA TotalPrep Amplification kit (cat no IL1791, http://www.ambion.com/techlib/prot/fm_IL1791.pdf)
|
| Label |
biotin
|
| Label protocol |
Biotinylate RNA was prepared using Ambion's Illumina RNA TotalPrep Amplification kit (cat no IL1791, http://www.ambion.com/techlib/prot/fm_IL1791.pdf)
|
| |
|
| Hybridization protocol |
1.50 ug of mRNA was hybridized to Illumina’s Sentrix Human-6 Expression BeadChips (cat no BD-25-101) at 55 C overnight (18,5 h) according to Illumina BeadStation 500X –protocol, Revision F.
|
| Scan protocol |
Hybridization was detected with 1 ug/ml Cyanine3-streptavidine, Amersham Biosciences (cat no 146065). Chips were scanned with Illumina BeadArray Reader PMT 625 and the numerical results were extracted with Bead Studio v1.5.1.34 without any normalization or background subtraction.
|
| Description |
Patient7C
|
| Data processing |
Data was normalized using the rank invariant method.
|
| |
|
| Submission date |
Oct 15, 2009 |
| Last update date |
Oct 15, 2009 |
| Contact name |
Mikael Benson |
| E-mail(s) |
mikael.benson@liu.se
|
| Phone |
+46 31 343 5162
|
| Fax |
+46 31 848952
|
| Organization name |
Queen Silvia Children's Hospital
|
| Department |
Paediatrics
|
| Street address |
Smorslottsgatan
|
| City |
Gothenburg |
| ZIP/Postal code |
416 85 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL2507 |
| Series (1) |
| GSE18574 |
Allergen-challenged CD4+ cells from patients with seasonal allergic rhinitis |
|
