|
| Status |
Public on Mar 28, 2013 |
| Title |
Donor-2, B cells (CD19+CD20+) |
| Sample type |
RNA |
| |
|
| Source name |
B cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: B cells
markers: CD19+CD20+
|
| Treatment protocol |
To examine gene expression in purified cellular fractions, normal human blood was collected from 2 donors as per institutional policy. The granulocyte (CD15+), monocyte (CD14+), T cell (CD3+), B cell (Non-PC gated, CD19+), and PC (CD27++CD38++) fractions from peripheral blood were separated. White blood cells were washed with FACS buffer (PBS + 0.5%BSA + 2mM EDTA (Gibco)) and incubated with 20% heat-inactivated FBS for 10-15 minutes on ice. The following mAbs were added directly to the cells: CD15 (HI98); CD14 (M5E2), CD3 (UCHT1), CD27 (M-T271), CD38 (HB7), and DAPI (Molecular probes). Cells were sorted on a Becton Dickinson FACS Aria II flow cytometer. All sorted fractions were collected in FACS buffer, centrifuged, and the resulting cell pellet was suspended in RNA lysis buffer (Ambion).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Zymo RNA MicroPrep kit extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Ambion MessageAmp II-Biotin Enhanced Single Round aRNA Amplification protocol from 75 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on HGU133 Plus 2.0 Array. GeneChips were washed and stained in an Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.
|
| Description |
HGU133 Plus array data
|
| Data processing |
The data was analyzed in R.
|
| |
|
| Submission date |
Mar 27, 2013 |
| Last update date |
Sep 01, 2016 |
| Contact name |
Christopher Allen Morehouse |
| Organization name |
Medimmune LLC
|
| Street address |
One Medimmune Way
|
| City |
Gaithersburg |
| State/province |
MD |
| ZIP/Postal code |
20878 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (2) |
| GSE45535 |
The Plasma Cell Signature in Autoimmune Disease (I) |
| GSE45537 |
The Plasma Cell Signature in Autoimmune Disease |
|
| Relations |
| Reanalyzed by |
GSE86362 |
