|
| Status |
Public on Aug 10, 2016 |
| Title |
M0-1 |
| Sample type |
RNA |
| |
|
| Source name |
Macrophage
|
| Organism |
Homo sapiens |
| Characteristics |
cell population: CD14+ monocytes
macrophage subtype: M0
|
| Treatment protocol |
CD14+ monocytes from human peripheral blood mononuclear cells were cultured with GM-CSF or M-CSF for seven days, and then stimulated for 24 h with LPS+ IFN-γ, IFN-γ, IL-4, IL-1β, and IL-10.
|
| Growth protocol |
Human Macrophages were cultured in RPMI1640 medium under 5% CO2 condition.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using the RNeasy Mini Kit (Qiagen), quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1 ug RNA using the Low Input Quick Amp Labeling kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA).
|
| |
|
| Hybridization protocol |
1.65 ug of Cy3-labelled cRNA (specific activity > 21pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Human Whole Genome Ver. 2.0 arrays (G4845A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner using one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green PMT is set to 100%).
|
| Description |
CD14+ monocytes were cultured with M-CSF for seven days, and then incubated for 24 h without additional factors
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5.1 (Agilent) using default parameters
|
| |
|
| Submission date |
Aug 09, 2016 |
| Last update date |
Aug 10, 2016 |
| Contact name |
Kyoko Yamashiro |
| E-mail(s) |
yamashiro.kyoko.r2@asubio.co.jp
|
| Organization name |
Asubio pharma co.,LTD.
|
| Street address |
6-4-3, inatojima-Minamimachi,
|
| City |
Chui-ku,Kobe |
| ZIP/Postal code |
650-0047 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13497 |
| Series (1) |
| GSE85346 |
Gene Expression Analysis in Different Macrophage Phenotypes |
|
