|
| Status |
Public on May 25, 2011 |
| Title |
BCTNN_BMP2_1of2 |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
beta-catenin ChIP DNA from BMP2 treated HPAEC
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: human pulmonary artery endothelial cells
chip antibody: beta-catenin
|
| Treatment protocol |
Cells were treated with recombinant human BMP2 (Sigma-Aldrich) for 4 hours before harvesting.
|
| Growth protocol |
HPAECs from large vessels (ScienCell) were grown in commercial EC media supplied by ScienCell (Cat: 1001) and used at passages 4-8.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
ChIP was performed on 10x10^6 HPAECs with anti-ppar-gamma (Santa Cruz; sc-7273X) and anti-beta-catenin (UPSTATE; Cat 06-734) antibodies using the Farnham ChIP protocol. The immunoprecipitated DNA was amplified using the Whole Genome Amplification Kit (Sigma).
|
| Label |
Cy5
|
| Label protocol |
Input DNA was labeled with Cy3 and ChIP DNA was labeled with Cy5 using the Standard NimbleGen ChIP labeling protocol (1 ug of amplified ChIP DNA, direct incorporation of Cy-labeled primers with Klenow extension).
|
| |
|
| Channel 2 |
| Source name |
Input DNA from BMP2 treated HPAEC
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: human pulmonary artery endothelial cells
chip antibody: none, input DNA
|
| Treatment protocol |
Cells were treated with recombinant human BMP2 (Sigma-Aldrich) for 4 hours before harvesting.
|
| Growth protocol |
HPAECs from large vessels (ScienCell) were grown in commercial EC media supplied by ScienCell (Cat: 1001) and used at passages 4-8.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
ChIP was performed on 10x10^6 HPAECs with anti-ppar-gamma (Santa Cruz; sc-7273X) and anti-beta-catenin (UPSTATE; Cat 06-734) antibodies using the Farnham ChIP protocol. The immunoprecipitated DNA was amplified using the Whole Genome Amplification Kit (Sigma).
|
| Label |
Cy3
|
| Label protocol |
Input DNA was labeled with Cy3 and ChIP DNA was labeled with Cy5 using the Standard NimbleGen ChIP labeling protocol (1 ug of amplified ChIP DNA, direct incorporation of Cy-labeled primers with Klenow extension).
|
| |
|
| |
| Hybridization protocol |
Standard NimbleGen hybridization kits and protocols were used and hybridization was performed using a NimbleGen 12 slide hybridization system.
|
| Scan protocol |
Arrays were scanned with Axon scanner at 5 um resolution according to standard NimbleGen protocols.
|
| Description |
HPAEC ChIP-chip anti-beta-catenin BMP2 treated 1of2
|
| Data processing |
Arrays were processed using Nimblegen's standard protocol for Nimblescan 2.5 ChIP data extraction. Log2 ChIP/input ratios were created from the two raw pair files.
|
| |
|
| Submission date |
May 23, 2011 |
| Last update date |
May 25, 2011 |
| Contact name |
Molong Li |
| E-mail(s) |
molong.li@jhmi.edu
|
| Organization name |
Johns Hopkins University
|
| Department |
School of Medicine
|
| Street address |
733 N. Broadway
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL6325 |
| Series (1) |
| GSE29489 |
ChIP-chip of BMP2 or control treated human pulmonary artery endothelial cells with anti-beta-catenin or anti-ppar-gamma antibodies on promoter regions |
|
