|
| Status |
Public on May 09, 2014 |
| Title |
Ly-6B+Ly-6G- bone marrow monocyte_biological rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
bone marrow monocyte, naive
|
| Organism |
Mus musculus |
| Characteristics |
gender: Female
strain: C57BL/6
tissue: Bone marrow
age: 7-8 weeks
|
| Treatment protocol |
Cells were blocked with 5% heated-rabbit serum, 0.5%BSA in PBS, 5mM EDTA (on ice) and then stained by the addition of fluorescently-labeled antibodies for 1 hour on ice. Free-antibody was removed by 2 washes with 0.5% BSA in PBS with 5mM EDTA before purifying the target populations on a MoFlo cell sorter. RNA was then extracted from purified cells.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using TriZol reagent performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
100ng RNA was amplified and labeled using the GeneChip® Two-Cycle Target Labeling and Control Reagents from Affymetrix.
|
| |
|
| Hybridization protocol |
15μg fragmented cDNA was then hybridised overnight to the Affymetrix MOE430 2.0 GeneChips
|
| Scan protocol |
GeneChips were scanned using the Agilent Gene Array Scanner controlled by GCOS 1.2
|
| Description |
Bones were flushed with ice-cold PBS, 0.5% BSA, 5mM EDTA and marrow was passed through a 40um cell strainer. Red blood cells were lysed with ACK buffer at room temperature and then cells were stained via the common protocol.
|
| Data processing |
The data were analyzed with Affymetrix Expression Console 1.2 using the MAS 5.0 algorithm and global scaling (TGT 100) as the normalization method.
|
| |
|
| Submission date |
Apr 14, 2011 |
| Last update date |
May 09, 2014 |
| Contact name |
Peter Giles |
| E-mail(s) |
GilesPJ@Cardiff.ac.uk
|
| Organization name |
Cardiff University
|
| Street address |
Heath Park
|
| City |
CARDIFF |
| ZIP/Postal code |
CF14 4XN |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE28621 |
Transcriptional profiles of macrophages in resolving inflammation |
|
