|
| Status |
Public on Apr 01, 2008 |
| Title |
unstimulated(confluent)-1 |
| Sample type |
RNA |
| |
|
| Source name |
human umbilical vein endothelial cells
|
| Organism |
Homo sapiens |
| Characteristics |
unstimulated confluent HUVECs
|
| Treatment protocol |
no treatment
|
| Growth protocol |
HUMEDIA; confluent growth
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNAs were extracted using Isogen (Nippon Gene, Osaka, Japan). 3 micrograms of total RNA was reverse transcribed using the SuperScript III system for cDNA synthesis (Invitrogen) according to the protocol recommended by Affymetrix. The oligonucleotide used for priming was 5'-ggccagtgaattgtaatacgactcactatagggaggcgg-(t)24-3' (Invitrogen). Double-stranded cDNA was cleaned by phenol:chloroform extraction and the aqueous phase removed by centrifugation through Phase-lock Gel (Eppendorf).
|
| Label |
biotin
|
| Label protocol |
In vitro transcription was performed on 1 microgram of cDNA using the Enzo BioArray High Yield RNA transcript labelling kit (Enzo Diagnostics). The cRNA was cleaned using RNAeasy clean-up columns (Qiagen). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.
|
| |
|
| Hybridization protocol |
10 micrograms of fragmented cRNA were hybridised (45degree CelsiusC, 16 hours). Hybridization was controled by use of the GeneChip Eukaryotic Hybridization Control Kit (Affymetrix). Washing and staining was performed in a Fluidics Station 450 (Affymetrix) using the protocol EukGE-WS2v4.
|
| Scan protocol |
Scanning was performed in an Affymetrix GeneChip scanner 3000 7G.
|
| Description |
Unstimulated HUVEC under confluent condition
|
| Data processing |
Affymetrix Microarray Suite v5.0 with target intensity=100
|
| |
|
| Submission date |
Nov 24, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
NAOKI MOCHIZUKI |
| E-mail(s) |
nmochizu@ri.ncvc.go.jp
|
| Phone |
+81-6-6833-5012
|
| Fax |
+81-6-6835-5461
|
| Organization name |
Natl. Cardiovasc. Ctr. Res. Inst
|
| Department |
Structural Analysis
|
| Street address |
5-7-1 Fujishirodai, Suita
|
| City |
Osaka |
| ZIP/Postal code |
565-8565 |
| Country |
Japan |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE9677 |
Gene expression profile in HUVECs before and after Angiopoietin stimulation |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE86362 |
| Reanalyzed by |
GSE119087 |
