|
| Status |
Public on Sep 22, 2009 |
| Title |
LC from biological replicate C57BL/6_2 |
| Sample type |
RNA |
| |
|
| Source name |
purified Langerhans cells from ear epidermis, isolated after 16 hours of ex vivo cultivation
|
| Organism |
Mus musculus |
| Characteristics |
C57BL/6 mice, 6-8 week old female mice
|
| Treatment protocol |
no treatment of mice
|
| Growth protocol |
epidermal sheets, cultivated for 16 hours, then isolation of LC by BSA gradient and FACS sorting
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with Trizol™ (Invitrogen) according to the manufacturers’ instructions. For microarray analysis mRNA was amplified prior to chip-hybridization using the MessageAmp™ Kit of Ambion (Woodward St. Austin, USA)
|
| Label |
biotin
|
| Label protocol |
RNA was biotinylated (Enzo Bio Array™HighYield™ RNA transcript labeling kit (Affymetrix, High Wycombe, UK))
|
| |
|
| Hybridization protocol |
standard Affymetrix protocol
|
| Scan protocol |
standard Affymetrix protocol
|
| Description |
For cell culturing of epidermal cells, an additional digestion with DNAse I (15 min, 130 U/ml) was included. The resulting single cell suspension was cultured in complete medium for 16 hours at a density of 1.5x106 cells/ml in 6-well plates. For sorting, epidermal cells were centrifuged on dense BSA or OptiPrep® (Sigma). The low-density fraction contained 20-35 % LC, which were enriched by FACS sorting of vital MHC-II+ cells using a FACSCalibur (Becton Dickinson, Heidelberg, Germany) to a purity >95 %. Sort purity was verified by re-analysis of samples.
|
| Data processing |
The resulting cel files were analyzed with the bioconductor affy package using the RMA (robust microarray analysis) algorithm.
|
| |
|
| Submission date |
Oct 08, 2008 |
| Last update date |
Sep 22, 2009 |
| Contact name |
Charlotte Esser |
| E-mail(s) |
chesser@uni-duesseldorf.de
|
| Organization name |
IUF
|
| Department |
Molecular Immunology
|
| Street address |
Auf´m Hennekamp 50
|
| City |
Dusseldorf |
| ZIP/Postal code |
40225 |
| Country |
Germany |
| |
|
| Platform ID |
GPL339 |
| Series (1) |
| GSE13130 |
Langerhans cells from aryl hydrocarbon receptor deficient mice |
|
