|
| Status |
Public on May 28, 2014 |
| Title |
MoDC Young_2 |
| Sample type |
RNA |
| |
|
| Source name |
MoDC from age_25 donor
|
| Organism |
Homo sapiens |
| Characteristics |
individual: young donor
age: 25
cell type: monocyte derived dendritic cells (MoDC)
|
| Treatment protocol |
Purified CD14+ monocytes were cultured in RPMI 1640 10% fetal bovine serum. Supplemented with human rGM-CSF,50ng/ml and 10 ng/ml human rIL-4.
|
| Growth protocol |
To induce differentiation of DCs, purified CD14+ monocytes were cultured in RPMI 1640 10% fetal bovine serum. Supplemented with human rGM-CSF,50ng/ml and 10 ng/ml human rIL-4 in 37C incubator with 5% CO2 for 6 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from MoDCs using the TRI Reagent kit (Molecular Research Center Inc., Cincinnati, OH, USA), following the manufacturer’s protocol.
|
| Label |
Biotin
|
| Label protocol |
The single-stranded cRNA sample is fragmented (WT Terminal Labeling Kit, Affymetrix, Inc, Santa Clara, CA) to an average strand length of 70 bases (range 40-70bp) following prescribed protocols (Affymetrix GeneChip WT Sense Target Labeling Assay Manual). The fragmented single-stranded DNA is subsequently labeled with recombinant terminal deoxynucleotidyl transferase (TdT) and the Affymetrix proprietary DNA Labeling Reagent that is covalently linked to biotin.
|
| |
|
| Hybridization protocol |
2 ug of fragmented target single-stranded cRNA was hybridized at 45c with rotation for 17 hours (Affymetrix GeneChip® Hybridization Oven 640) to probe sets present on an Affymetrix array. The GeneChip® arrays were washed and then stained (SAPE, streptavidin-phycoerythrin) on an Affymetrix Fludics Station 450 (Fluidics protocol FS450_007).
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 7G and Command Console Software v. 3.2.3.to produce cell intensity files (CEL). CEL were analyzed with Microarray Suite 5.0
|
| Description |
Gene expression data in MoDC from young donor
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100.
|
| |
|
| Submission date |
May 28, 2014 |
| Last update date |
May 28, 2014 |
| Contact name |
Sudhir Gupta |
| Organization name |
University of California, Irvine
|
| Department |
Medicine
|
| Lab |
Immunology
|
| Street address |
Med Sci Buld I, C240
|
| City |
Irvine |
| State/province |
CA |
| ZIP/Postal code |
92697 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE58015 |
Gene Expression Data in Human Monocyte-derived Dendritic Cell (MoDC) from Young and Aged Donors |
|
