|
| Status |
Public on Sep 30, 2015 |
| Title |
non-classical monocytes |
| Sample type |
RNA |
| |
|
| Source name |
non-classical monocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD14+CD16++
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Human monocytes were isolated from buffy coats from healthy donors, and sorted by FACS analysis using specific antibodies. Human advanced atherosclerotic plaques were collected during carotid endarterectomy. Aa. mammariae internae excised from patients undergoing elective by-pass surgery. Total RNA was isolated using the TRIzol kit (Invitrogen). miR was purified using the mirVanaTM miR Isolation Kit (Ambion).
|
| Label |
cy3
|
| Label protocol |
RNA from plaque tissue and mamaria were labeled using the mirVanaTM miR Labeling Kit (Ambion) and fluorescent Cy3 (Molecular Probes). RNA from human monocytes were labeled with GenoExplorer™ miRNA Labeling Kit.
|
| |
|
| Hybridization protocol |
Probes generated from plaque tissue and mamaria were hybridized to the Ambion mirVanaTM miR Bioarray (1566 v.1, www.ambion.com/techlib/prot) according to manufactory instruction. Probes generated from human monocytes were hybridized to GenoExplorer human microRNA array by BIOCAT service also according to manufactory instruction.
|
| Scan protocol |
Scanning of the Ambion mirVana Bioarrays was performed using a dual-laser scanner Affymetrix 428 (affymetrx, CA) at various amplification settings (gains) above and below saturation of the most intensely fluorescent spots on each array. GenoExplorer human microRNA arrays scanning were provided by BIOCAT service.
|
| Data processing |
ImaGene 5.5.4 (BioDiscovery, CA) software was used to quantify the scanned Ambion mirVana Bioarray images. GenoExplorer human microRNA arrays were quatified BIOCAT service. Median pixel intensities of the spots were taken and normalized based on control spot intensities. Spots with intensities lower than negative control intensities were defined as 0, spots with intensities higher than negative controls get a relative expression value by comparing to positive control intensities.
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| |
|
| Submission date |
Sep 12, 2011 |
| Last update date |
Sep 30, 2015 |
| Contact name |
Lin Gan |
| Organization name |
University Hospital Aachen
|
| Department |
IZKF
|
| Lab |
Genomics facility
|
| Street address |
Pauwelsstr. 30
|
| City |
Aachen |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10860 |
| Series (1) |
| GSE32114 |
microRNA expression signatures and parallels between monocyte subsets and atherosclerotic plaque in humans |
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