|
| Status |
Public on Oct 01, 2008 |
| Title |
vehicle_6h_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Raw264.7 macrophage cells as vehicle controls
|
| Organism |
Mus musculus |
| Characteristics |
macrophage-like cell line
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were treated with LPS in the presence or absence of Paeony root components for 6h. Total RNA was extracted using Trizol reagent.
|
| Label |
DIG-UTP
|
| Label protocol |
Digoxigenin (DIG)-labeled cRNA was generated from the total RNA by using the chemiluminescent RT-IVT labeling kit (Applied Biosystems) as per the manufacturer’s instruction. 2 ug of total RNA was reverse transcribed for 2 h at 42℃. The second strand DNA was synthesized for 2 h at 16℃. The double stranded cDNA was purified and mixed with DIG-UTP (Roche), IVT enzyme mix, and IVT buffer mix. IVT reaction was performed for 9 h at 37℃.
|
| |
|
| Hybridization protocol |
The microarray chip was pre-hybridized with hybridization buffer (Applied Biosystems) for 1 h at 55℃. cRNA fragmentation was for 30 min at 60℃. The fragmented cRNA was added to prehybridized chip and the chip was incubated for 16 h at 55℃ with constant rocking at 100 rpm.
|
| Scan protocol |
The microarray chip was scanned with Applied Biosystems 1700 Chemiluminescent Microarray Analyzer.
|
| Description |
The RAW264.7 macrophages were either treated with DMSO or with LPS in the presence or absence of albiflorin, paeoniflorin, or paeonol.
|
| Data processing |
The raw data were normalized using median shifting and lowess regression normalization methods by avadis software (India). Log-tansformation algorithm was used.
|
| |
|
| Submission date |
Nov 16, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Hoog-Hee Kim |
| E-mail(s) |
hhbkim@snu.ac.kr
|
| Organization name |
Seoul National University
|
| Department |
Cell and Developmental Biology
|
| Street address |
28 Yeongon-Dong, Chongno-Gu,
|
| City |
Seoul |
| ZIP/Postal code |
110-749 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL2995 |
| Series (1) |
| GSE9632 |
Gene expression profiles of RAW264.7 macrophage cells treated with Paeony root-derived compounds |
|
