|
| Status |
Public on Nov 07, 2011 |
| Title |
macrophage_untreated_03 |
| Sample type |
RNA |
| |
|
| Source name |
human peripheral blood monocytes, M-CSF differntiated to macrophages
|
| Organism |
Homo sapiens |
| Characteristics |
Species: Human, healthy; Cell Type: Macrophages; M-CSF differntiated for 4 days; Pooled Material;
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cells according to the manufacturer’s instructions using the RNeasy Protect Midi Kit (Qiagen, Hilden, Germany). Purity and integrity of the RNA was assessed on the Agilent 2100 bioanalyzer with the RNA 6000 Nano LabChip reagent set (Agilent Technologies, Böblingen, Germany). The RNA was quantified spectrophotometrically and then stored at −80°C.
|
| Label |
biotin
|
| Label protocol |
Following phenol-chloroform extraction, cDNA was used for in vitro transcription reaction using the T7 BioArray High Yield RNA Transcript Labeling Kit (Enzo Diagnostics, Farmingdale, NY) to produce biotinylated cRNA. Thereafter, the labeled cRNA was purified using RNeasy Mini Kit columns (Qiagen) and fragmented by incubation at 94°C for 30 minutes.
|
| |
|
| Hybridization protocol |
All microarrays were processed in an Affymetrix GeneChip Fluidics Station 400. Following staining with streptavidin-conjugated phycoerythrin and washing cycles.
|
| Scan protocol |
Microarrays were scanned using the GeneArray Scanner (Agilent Technologies).
|
| Description |
Differentiation was confirmed by phase contrast microscopy.
The fragmentation was checked by microcapillary electrophoresis on an Agilent 2100 bioanalyzer, and 20 μg of biotinylated cRNAs was hybridized to U133A GeneChips for 16 hours at 45°C with constant rotation.
|
| Data processing |
Expression signals for each transcript and comparisons between different samples were calculated with the Affymetrix GeneChip software MAS5.0.
|
| |
|
| Submission date |
Dec 06, 2007 |
| Last update date |
Nov 07, 2011 |
| Contact name |
Dominik Lutter |
| E-mail(s) |
[email protected]
|
| Organization name |
Helmholtz Zentrum München
|
| Department |
Institute for Diabetes and Obesity
|
| Street address |
Ingolstädter Landstraße 1
|
| City |
Neuherberg |
| ZIP/Postal code |
D-85764 |
| Country |
Germany |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE9801 |
Human Monocytes to M-CSF differentiated Macrophages |
|
