|
| Status |
Public on Nov 01, 2013 |
| Title |
MCF7 GF200 001 |
| Sample type |
protein |
| |
|
| Source name |
conditioned media from MCF7 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF7
cell type: breast cancer
|
| Treatment protocol |
MCF7 were treated with 5 ng/mL TGF-β , 10 ng/mL TNF-α(PeproTech, Rocky Hill, NJ) in 10%FBS DMEM for 2 weeks or given prolonged mammosphere culture for 5 weeks as previously described (Guttilla et al., 2012).
|
| Growth protocol |
MDA-MB-23 and MCF-7 breast cancer cells were obtained from American Type Culture Collection (ATCC) and grown according to standard protocols.
|
| Extracted molecule |
protein |
| Extraction protocol |
Afterward, cells were washed in PBS and grown in adherent condition with cytokine/serum-free media which were harvested 24 hours later. The conditioned media was centrifuged at 4 。C for 20 min and the cell-free supernatant was collected.
|
| Label |
biotin
|
| Label protocol |
Samples were labeled according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Human Cytokine Antibody Arrays V kit (Raybiotech) was used according to the manufacturer’s instructions. Briefly, the arrays were blocked, incubated with 100 μL of CM overnight, incubated with biotin-conjugated antibodies (1/250) for 2 hours and with HRP-linked secondary antibody (1/1000) overnight. The membranes were incubated with chemiluminescent substrate and exposed to x-ray film for 15 minutes before development.
|
| Scan protocol |
Quantitative array analysis was performed using Array Vision Evaluation 8.0 (GE Healthcare Life Science)
|
| Data processing |
To obtain relative expression, signal intensity was subtracted for negative controls and normalized to internal positive. The intensities of signals can be quantified by densitometry.
|
| |
|
| Submission date |
Oct 31, 2013 |
| Last update date |
Nov 01, 2013 |
| Contact name |
Shicheng Su |
| E-mail(s) |
seasonso@163.com
|
| Phone |
84-02-87332022
|
| Organization name |
Sun-Yat-Sen Memorial Hospital
|
| Street address |
107 Yanjiang West Road
|
| City |
Guangzhou |
| ZIP/Postal code |
510120 |
| Country |
China |
| |
|
| Platform ID |
GPL17868 |
| Series (1) |
| GSE51938 |
Analysis of conditioned media cytokine levels of breast cancer cell lines |
|
