|
| Status |
Public on Feb 05, 2008 |
| Title |
Free mRNP of J774-LPS 4hr-rep3 |
| Sample type |
RNA |
| |
|
| Source name |
macrophage-like J774 cells, free mRNP, LPS 4hr
|
| Organism |
Mus musculus |
| Characteristics |
Strain:Balb/c Tissue:blood macrophage-like cell
|
| Biomaterial provider |
RIKEN bioresource center
|
| Treatment protocol |
100ng/ml of Lipopolysaccharide from Escherichia coli (sero type 055:B5)
|
| Growth protocol |
RPMI1649 10%FCS 37C, 5%CO2 passage:twice/week
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA was extracted with TRIzol reagent(Invitrogen), and subsequently poly(A) RNA was isolated using microMACS kit(Militenyi). Qulity of poly(A) RNA was checked using a bioanalyzer (Agilent).
|
| Label |
biotin
|
| Label protocol |
Labeling was performed using One-cycle target labeling kit (Affymetrix) as manufacture's indications.
|
| |
|
| Hybridization protocol |
Twenty micro gram of labeled sample was used. Hybridization was performed at 45C for 16hr with continuous rotation.
|
| Scan protocol |
Washing and streptavidin-phycoerythrin staining was done using a GeneChip Fluidics Station(Affymetrix), then scanning was performed using a GeneChip scanner 3000(Affymetrix) as the default setting.
|
| Description |
Free mRNP preparation: The cells were lysed with a solution containing 0.5%NP-40.Post-mitochondrial fraction was applied onto 15-50% sucrose gradient, and subsequently centrifuged using SW41Ti (Beckman Coulter) at 39krpm. The gradient was divided into 20 fractions from the surface. The fraction 1-5 were collected, and subjected to total RNA extraction.
|
| Data processing |
The value was calculated by MAS5 method using a GCOS software package(Affymetrix)
|
| |
|
| Submission date |
Jan 25, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Hiroshi Kitamura |
| E-mail(s) |
ktmr@rakuno.ac.jp
|
| Phone |
81-11-388-4781
|
| Organization name |
Rakuno Gakuen University
|
| Department |
School of Veterinary Medicine
|
| Lab |
Laboratory for Veterinary Physiology
|
| Street address |
582 Midorimachi, Bunkyodai
|
| City |
Ebetsu |
| ZIP/Postal code |
069-8501 |
| Country |
Japan |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE4288 |
Polysomal, free mRNP, and total cellular mRNA of J774.1 before and after LPS stimulation |
|
| Relations |
| Reanalyzed by |
GSE119085 |
