|
| Status |
Public on Oct 02, 2010 |
| Title |
LYVE-1 negative macrophages, rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
WT mouse skin LYVE-1 negative macrophages
|
| Organism |
Mus musculus |
| Characteristics |
age: embryonic day 15.5
|
| Treatment protocol |
Embryonic skin was dissected in HHF (HBBS with HEPES and FBS) and digested in 5 ml of digestion buffer (DMEM with FBS with Collagenase II (Worthington), Collagenase IV (Worthington) and Deoxyribonuclease I (Worthington)) per 5-8 embryos. Digestion was performed at 37°C for 30 min. Macrophages were isolated on the basis of F4/80 expression using a magnetic cell separation procedure (MACS®) (Miltenyi Biotec) according to manufacturers instructions. F4/80-positive macrophages were separated on the basis of LYVE-1 expression using a MACS® procedure (Miltenyi Biotec) according to manufacturers instructions.
|
| Growth protocol |
Wild-type embryos were collected at embryonic day 15.5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of RNA was performed using RNAqueous (Ambion) according to manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix Genearray protocol from 100ng total RNA (Affymetrix GeneChip Whole Transcript [WT] Sense Target Labeling Assay, rev 4, Affymetrix 2005-2007).
|
| |
|
| Hybridization protocol |
Following fragmentation, 2.5 ug of cDNA were hybridized for 16 hr at 45C on GeneChip Mouse Gene 1.0 ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G and GCOS software v1.4
|
| Description |
Gene expression data from embryonic skin macrophages
|
| Data processing |
The data were analyzed with Partek Genomics Suite v6.3 (RMA) using quantile normalization and median polish probeset summarization.
|
| |
|
| Submission date |
Oct 01, 2010 |
| Last update date |
Oct 01, 2010 |
| Contact name |
Emma Gordon |
| E-mail(s) |
[email protected]
|
| Phone |
61 8 8222 3526
|
| Organization name |
SA Pathology
|
| Department |
Haematology
|
| Lab |
Lymphatic Development Laboratory
|
| Street address |
Frome Road
|
| City |
Adelaide |
| State/province |
South Australia |
| ZIP/Postal code |
5000 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE24492 |
Investigation of the gene expression profile of LYVE-1-positive macrophages |
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