|
| Status |
Public on Jan 14, 2014 |
| Title |
BC4869 CD40-activated B cell |
| Sample type |
RNA |
| |
|
| Source name |
peripheral blood
|
| Organism |
Homo sapiens |
| Characteristics |
donor id: BC4869
tissue: peripheral blood
cell type: CD19-positive B cells
stimulated with: CD40
cell status: CD40-activated
|
| Treatment protocol |
Human B cells were purified from buffy coats using CD19 MACS.
|
| Growth protocol |
For CD40-activation the purified CD19-positive B cells were stimulated via CD40 by means of NIH3T3 cells transfected with CD40L in 6-well plates in medium containing 45% Dulbecco modified Eagle medium supplemented with 10% fetal calf serum, 2 mM glutamine, and 15 ug/mL gentamicin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Immature monocyte-derived DC were harvested on day 7, lyzed in TRIzol and stored at 80°C until further processing. RNA was extracted from cell lyzates with chloroform/isopropanol method, and was clean-uped in silica columns (Qiagen). The quality of the isolated RNA was determined by measuring the A260/280 ratio and the integrity of the ribosomal 28s and 18s bands were determined by agarose-gel electrophoresis.
|
| Label |
Biotin
|
| Label protocol |
Synthesis of biotin labeled cRNA was performed with BioArray HighYield RNA TranscriptLabeling Kit (ENZO) following standard procedures.
|
| |
|
| Hybridization protocol |
Hybridization and washing was performed following standard procedures for Affymetrix HG-U133A arrays (www.affymetrix.com) on a GeneChip Fluidic station 400, using the EukGe-WS2v4 protocol.
|
| Scan protocol |
Scanning was performed twice with filter set at 570 nm on an Agilent gene array scanner (Affymetrix).
|
| Description |
BC4869_CD40B_U133A
|
| Data processing |
MAS5.0 (Affymetrix) was used for raw data measurement and quality control. dChip 2006 was used for data normalization, quality control and data analysis.
|
| |
|
| Submission date |
Jan 13, 2014 |
| Last update date |
Sep 01, 2016 |
| Contact name |
Alexander Shimabukuro-Vornhagen |
| Organization name |
University Hospital of Cologne
|
| Department |
Department I of Internal Medicine
|
| Lab |
Cologne Interventional Immunology
|
| Street address |
Kerpener Str. 62
|
| City |
Cologne |
| ZIP/Postal code |
50674 |
| Country |
Germany |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE54017 |
CD40-activation of human B cells |
|
| Relations |
| Reanalyzed by |
GSE86363 |
