|
| Status |
Public on Oct 22, 2021 |
| Title |
only LPS (4μg/ml) treatment biol rep 3 tech rep 3 |
| Sample type |
RNA |
| |
|
| Source name |
U937 cell line (human histiocytic lymphoma cell)
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: U937
treatment: only LPS (4μg/ml)
|
| Treatment protocol |
Human macrophage cells were untreatment, LPS (4μg/ml) only, preteated PBD150 inhibitor (10μM) for 2 hrs before the addition LPS (4μg/ml) treatment, the incubation time of the culture was 48 hours.
|
| Growth protocol |
The U937 cells were maintained at 2-3 × 106 cells/ml in a RPMI 1640 medium supplemented with 10% FBS, the incubation time of the culture was 48 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction by column based extraction kit (RiboPure-Blood, Ambion). RNA purity was checked by optical density of NanoDrop ND-1000 and agarose electrophoresis. RNA integrity was measured by Agilent RNA 6000 Nano Assay (RIN>8).
|
| Label |
Cy5
|
| Label protocol |
2.5 µg of total RNA was reverse-transcribed and amplified using MessageAmpTM aRNA Amplification Kit (Ambion). Indirect labeling the aa-UTP whereon by NHS-CyDye (Cy5, Amershan).
|
| |
|
| Hybridization protocol |
HOA v4.3 arrays were pre-heat at 60℃ for 10 mins, rehydrated by 100% ethanol following with deionized water. The slides were pre-hybridized with 5x SSPE, 0.1% SDS and 1% BSA at 42℃ for 2 hour. After the pre-hybridization, 5 µg Cy5-labeled aRNA was hybridize on HOA in the presentation of the Phalanx OneArray hybridization buffer.
|
| Scan protocol |
The arrays were scanned by Axon GenePix 4000B scanner (635nm power 100 PMT 600~630 ; 532nm power 10, PMT 460) and quantify the fluoresence intensity.
|
| Description |
Phalanx Biotech Group's Human OneArray contains 32,048 features, 30968 detection probes and 1097 control probes, spotted onto glass slides using a proprietary non-contact printing method.
|
| Data processing |
The raw data were adjust by Rosetta Resolver® error model calculation. The statistic values were calculated after the replicated probes queezing and median scaling normalization.
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| |
|
| Submission date |
Mar 26, 2010 |
| Last update date |
Oct 22, 2021 |
| Contact name |
Chen Yi Ling |
| Organization name |
Academia Sinica
|
| Department |
Institute of Biological Chemistry
|
| Lab |
501
|
| Street address |
128 Academia Road, Section 2
|
| City |
Taipei |
| ZIP/Postal code |
11529 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL6254 |
| Series (1) |
| GSE21082 |
Glutaminyl cyclase (QC) activity affects gene expression in macrophages during inflammation |
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