|
| Status |
Public on Aug 07, 2007 |
| Title |
NK_timepoint24hours |
| Sample type |
RNA |
| |
|
| Source name |
Natural killer cells
|
| Organism |
Homo sapiens |
| Characteristics |
Primary NK cells were isolated from healthy donors and peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Hypaque density gradient centrifugation at room temperature. Purified NK cells were cultured in RPMI 1640 medium (Life Technologies, New York, NY) supplemented with 10% fetal bovine serum, 200 ug/ml streptomycin and 200 IU/ml penicillin, and IL2 (100 IU/ml) for 24 hours at 37ºC and 5% CO2, before harvest and storage in Trizol at -80°C till RNA isolation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol and further purified with RNeasy Mini Columns (Qiagen, Valencia, CA) before aliquots were run in agarose gel electrophoresis and measured by spectrophotometer at 260 and 280 nm to assess the quality of the RNA.
|
| Label |
Biotin-pycoerythrin
|
| Label protocol |
http://www.affymetrix.com/products/reagents/specific/one_cycle_target_control.affx
|
| |
|
| Hybridization protocol |
http://www.affymetrix.com/products/reagents/specific/eukhyb.affx
|
| Scan protocol |
http://www.affymetrix.com/products/instruments/specific/scanner_3000.affx
|
| Description |
The isolation of RNA, hybridization and image processing was performed strictly according to the protocol set up by Affymetrix (Affymetrix Inc, CA). Briefly, extracted RNA (at least 2.6g used for each time point) was assessed by the Bio-analyzer 2100 (Agilent Inc, CA), and single round amplification protocol (one-cycle target labeling kit, Affymetrix Inc) was utilized for RNA amplification. Antisense biotin-labeled cRNA (10g), generated by in vitro transcription were hybridized and biotinylated probes hybridized to the array were detected using pycoerythrin labeled streptavidin. All arrays were scanned using the Gene-chip scanner 3000 (Affymetrix Inc, CA).
|
| Data processing |
The Affymetrix GeneChip® Operating Software (GCOS) was utilized for management, sharing, and analysis of data generated.
|
| |
|
| Submission date |
Jun 07, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Javeed Iqbal |
| E-mail(s) |
jiqbal@unmc.edu
|
| Phone |
402-559-7753
|
| Organization name |
University of Nebraska Medical Center
|
| Department |
Pathology and Microbiology
|
| Lab |
Center for Research in Leukemia and Lymphoma
|
| Street address |
40th street
|
| City |
omaha |
| State/province |
NE |
| ZIP/Postal code |
68195 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE8059 |
resting and IL2 activated human natural killer cells |
|
| Relations |
| Reanalyzed by |
GSE49910 |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE86362 |
| Reanalyzed by |
GSE119087 |
