|
| Status |
Public on Jun 14, 2024 |
| Title |
Control U937 cells treated LPS |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
U937 cells treated LPS
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: U937 (CRL-1593.2)
age: 37 years
sample type: Pleural effusion
gender: Male
|
| Treatment protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Growth protocol |
U937 cells were cultured in RPMI 1640 medium (Hyclone; SH30027.02) supplemented with 10% fetal bovine serum (Hyclone; SH30071.03), 2 mM L-glutamine (Simply; Cat. No CC515-0100), 100 units/mL Antibiotic-Antimycotic (Simply; Cat. No CC501-0100), and 1M HEPES (Simply; Cat. No CC519-0100) at 37℃ in a humidified atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The Low Input Quick Amp Labeling Kit, Two-Color generates fluorescent cRNA (complimentary RNA) with a sample input RNA range between 10 ng and 200 ng of total RNA or a minimum of 5 ng of poly A+ RNA for two-color processing. The method uses T7 RNA Polymerase Blend (red cap), simultaneously amplifying target material and incorporating Cyanine 3-CTP or Cyanine 5-CTP. Amplification is typically at least 100-fold from total RNA to cRNA with the use of this kit.
|
| Label |
Cy5
|
| Label protocol |
Oligoarray control targets and hybridization buffer (Agilent Two-color Microarray-BasedGene Expression Analysis V5) were added, and samples were applied to microarrays enclosed in Agilent microarray hybridization chambers. After hybridization, slides were washed sequentially
|
| |
|
| Channel 2 |
| Source name |
U937 cells treated LPS
|
| Organism |
Homo sapiens |
| Characteristics |
gender: Male
age: 37 years
sample type: Pleural effusion
|
| Treatment protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Growth protocol |
U937 cells were cultured in RPMI 1640 medium (Hyclone; SH30027.02) supplemented with 10% fetal bovine serum (Hyclone; SH30071.03), 2 mM L-glutamine (Simply; Cat. No CC515-0100), 100 units/mL Antibiotic-Antimycotic (Simply; Cat. No CC501-0100), and 1M HEPES (Simply; Cat. No CC519-0100) at 37℃ in a humidified atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The Low Input Quick Amp Labeling Kit, Two-Color generates fluorescent cRNA (complimentary RNA) with a sample input RNA range between 10 ng and 200 ng of total RNA or a minimum of 5 ng of poly A+ RNA for two-color processing. The method uses T7 RNA Polymerase Blend (red cap), simultaneously amplifying target material and incorporating Cyanine 3-CTP or Cyanine 5-CTP. Amplification is typically at least 100-fold from total RNA to cRNA with the use of this kit.
|
| Label |
Cy3
|
| Label protocol |
Oligoarray control targets and hybridization buffer (Agilent Two-color Microarray-BasedGene Expression Analysis V5) were added, and samples were applied to microarrays enclosed in Agilent microarray hybridization chambers. After hybridization, slides were washed sequentially
|
| |
|
| |
| Hybridization protocol |
Scanned on an Agilent Microarray scanner.
|
| Scan protocol |
Images were quantified using Agilent Feature Extraction Software (version A.10.5.1.1).
|
| Description |
Control U937 cells, treated with LPS, were collected after 72 hours.
|
| Data processing |
Agilent Feature Extraction Software (v 10.5.1.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Jun 13, 2024 |
| Last update date |
Jun 14, 2024 |
| Contact name |
Chang-Jung Chen |
| E-mail(s) |
openty2002@gmail.com
|
| Organization name |
National Cheng Kung University Medical College
|
| Street address |
Sheng-Li Road
|
| City |
Tainan |
| ZIP/Postal code |
70403 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE269815 |
U937 cells: Control treated LPS vs. Co-cultured Panc-1 cells |
|
